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1

Sidorova, Maria, Golo Kronenberg, Susann Matthes, et al. "Enduring Effects of Conditional Brain Serotonin Knockdown, Followed by Recovery, on Adult Rat Neurogenesis and Behavior." Cells 10, no. 11 (2021): 3240. http://dx.doi.org/10.3390/cells10113240.

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Serotonin (5-hydroxytryptamine, 5-HT) is a crucial signal in the neurogenic niche of the hippocampus, where it is involved in antidepressant action. Here, we utilized a new transgenic rat model (TetO-shTPH2), where brain 5-HT levels can be acutely altered based on doxycycline (Dox)-inducible shRNA-expression. On/off stimulations of 5-HT concentrations might uniquely mirror the clinical course of major depression (e.g., relapse after discontinuation of antidepressants) in humans. Specifically, we measured 5-HT levels, and 5-HT metabolite 5-HIAA, in various brain areas following acute tryptophan
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2

Berger, Stefan M., Iván Fernández-Lamo, Kai Schönig, et al. "Forebrain-specific, conditional silencing of Staufen2 alters synaptic plasticity, learning, and memory in rats." Genome Biology 18, no. 1 (2017): 222. https://doi.org/10.1186/s13059-017-1350-8.

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<strong>Background: </strong>Dendritic messenger RNA (mRNA) localization and subsequent local translation in dendrites critically contributes to synaptic plasticity and learning and memory. Little is known, however, about the contribution of RNA-binding proteins (RBPs) to these processes in vivo.<strong>Results: </strong>To delineate the role of the double-stranded RBP Staufen2 (Stau2), we generate a transgenic rat model, in which Stau2 expression is conditionally silenced by Cre-inducible expression of a microRNA (miRNA) targeting Stau2 mRNA in adult forebrain neurons. Known physiological mRN
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Zhang, Hui, Neil B. Sweezey, and Feige Kaplan. "LGL1 modulates proliferation, apoptosis, and migration of human fetal lung fibroblasts." American Journal of Physiology-Lung Cellular and Molecular Physiology 308, no. 4 (2015): L391—L402. http://dx.doi.org/10.1152/ajplung.00119.2014.

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Rapid growth and formation of new gas exchange units (alveogenesis) are hallmarks of the perinatal lung. Bronchopulmonary dysplasia (BPD), common in very premature infants, is characterized by premature arrest of alveogenesis. Mesenchymal cells (fibroblasts) regulate both lung branching and alveogenesis through mesenchymal-epithelial interactions. Temporal or spatial deficiency of late-gestation lung 1/cysteine-rich secretory protein LD2 (LGL1/CRISPLD2), expressed in and secreted by lung fibroblasts, can impair both lung branching and alveogenesis (LGL1 denotes late gestation lung 1 protein; L
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Flemming, Sven, Anny-Claude Luissint, Dennis H. M. Kusters, et al. "Desmocollin-2 promotes intestinal mucosal repair by controlling integrin-dependent cell adhesion and migration." Molecular Biology of the Cell 31, no. 6 (2020): 407–18. http://dx.doi.org/10.1091/mbc.e19-12-0692.

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We report a key role for Dsc2 in simple epithelial cell migration and mucosal wound healing in vivo using newly generated mice with inducible conditional knockdown of Dsc2 in intestinal epithelial cells ( Villin-CreERT2; Dsc2fl/fl).
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5

Ogawa, Yoshitaka, Kohei Nishimura, Keisuke Obara, and Takumi Kamura. "Development of AlissAID system targeting GFP or mCherry fusion protein." PLOS Genetics 19, no. 6 (2023): e1010731. http://dx.doi.org/10.1371/journal.pgen.1010731.

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Conditional control of target proteins using the auxin-inducible degron (AID) system provides a powerful tool for investigating protein function in eukaryotes. Here, we established an Affinity-linker based super-sensitive auxin-inducible degron (AlissAID) system in budding yeast by using a single domain antibody (a nanobody). In this system, target proteins fused with GFP or mCherry were degraded depending on a synthetic auxin, 5-Adamantyl-IAA (5-Ad-IAA). In AlissAID system, nanomolar concentration of 5-Ad-IAA induces target degradation, thus minimizing the side effects from chemical compounds
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kun, Zhang, Yang yuling, Wang dongchun, Xie bingbing, Li xiaoli та Xu bin. "HIF-1α Inhibition Sensitized Pituitary Adenoma Cells to Temozolomide by Regulating Presenilin 1 Expression and Autophagy". Technology in Cancer Research & Treatment 15, № 6 (2016): NP95—NP104. http://dx.doi.org/10.1177/1533034615618834.

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Pituitary adenomas usually develop temozolomide resistance, which could compromise the anticancer effects of temozolomide. Suppression of hypoxia-inducible factor 1α has been shown to sensitize glioblastoma cells to temozolomide treatment according to previous reports. However, whether and how the suppression of hypoxia-inducible factor 1α could sensitize pituitary adenomas to temozolomide treatment are still poorly understood. In the present study, using hypoxia-inducible factor 1α knockdown strategy, we demonstrated for the first time that hypoxia-inducible factor 1α knockdown could inhibit
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7

Makino‐Itou, Hatsune, Noriko Yamatani, Akemi Okubo, et al. "Establishment and characterization of mouse lines useful for endogenous protein degradation via an improved auxin‐inducible degron system (AID2)." Development, Growth & Differentiation 66, no. 7 (2024): 384–93. http://dx.doi.org/10.1111/dgd.12942.

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AbstractThe development of new technologies opens new avenues in the research field. Gene knockout is a key method for analyzing gene function in mice. Currently, conditional gene knockout strategies are employed to examine temporal and spatial gene function. However, phenotypes are sometimes not observed because of the time required for depletion due to the long half‐life of the target proteins. Protein knockdown using an improved auxin‐inducible degron system, AID2, overcomes such difficulties owing to rapid and efficient target depletion. We observed depletion of AID‐tagged proteins within
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8

Wiznerowicz, Maciej, and Didier Trono. "Conditional Suppression of Cellular Genes: Lentivirus Vector-Mediated Drug-Inducible RNA Interference." Journal of Virology 77, no. 16 (2003): 8957–51. http://dx.doi.org/10.1128/jvi.77.16.8957-8951.2003.

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ABSTRACT RNA interference has emerged as a powerful technique to downregulate the expression of specific genes in cells and in animals, thus opening new perspectives in fields ranging from developmental genetics to molecular therapeutics. Here, we describe a method that significantly expands the potential of RNA interference by permitting the conditional suppression of genes in mammalian cells. Within a lentivirus vector background, we subjected the polymerase III promoter-dependent production of small interfering RNAs to doxycycline-controllable transcriptional repression. The resulting syste
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9

Tsai, Steven C., David F. Chang, Chang-Mu Hong, et al. "Induced overexpression of OCT4A in human embryonic stem cells increases cloning efficiency." American Journal of Physiology-Cell Physiology 306, no. 12 (2014): C1108—C1118. http://dx.doi.org/10.1152/ajpcell.00205.2013.

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Our knowledge of the molecular mechanisms underlying human embryonic stem cell (hESC) self-renewal and differentiation is incomplete. The level of octamer-binding transcription factor 4 (Oct4), a critical regulator of pluripotency, is precisely controlled in mouse embryonic stem cells. However, studies of human OCT4 are often confounded by the presence of three isoforms and six expressed pseudogenes, which has complicated the interpretation of results. Using an inducible lentiviral overexpression and knockdown system to manipulate OCT4A above or below physiological levels, we specifically exam
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10

Weis, Sara M., Ssang-Taek Lim, Kimberly M. Lutu-Fuga, et al. "Compensatory role for Pyk2 during angiogenesis in adult mice lacking endothelial cell FAK." Journal of Cell Biology 181, no. 1 (2008): 43–50. http://dx.doi.org/10.1083/jcb.200710038.

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Focal adhesion kinase (FAK) plays a critical role during vascular development because knockout of FAK in endothelial cells (ECs) is embryonic lethal. Surprisingly, tamoxifen-inducible conditional knockout of FAK in adult blood vessels (inducible EC–specific FAK knockout [i-EC-FAK-KO]) produces no vascular phenotype, and these animals are capable of developing a robust growth factor–induced angiogenic response. Although angiogenesis in wild-type mice is suppressed by pharmacological inhibition of FAK, i-EC-FAK-KO mice are refractory to this treatment, which suggests that adult i-EC-FAK-KO mice
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11

Wey, Shiuan, Biquan Luo, Chun-Chih Tseng, et al. "Inducible knockout of GRP78/BiP in the hematopoietic system suppresses Pten-null leukemogenesis and AKT oncogenic signaling." Blood 119, no. 3 (2012): 817–25. http://dx.doi.org/10.1182/blood-2011-06-357384.

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Abstract Traditionally, GRP78 is regarded as protective against hypoxia and nutrient starvation prevalent in the microenvironment of solid tumors; thus, its role in the development of hematologic malignancies remains to be determined. To directly elucidate the requirement of GRP78 in leukemogenesis, we created a biallelic conditional knockout mouse model of GRP78 and PTEN in the hematopoietic system. Strikingly, heterozygous knockdown of GRP78 in PTEN null mice is sufficient to restore the hematopoietic stem cell population back to the normal percentage and suppress leukemic blast cell expansi
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12

Kotnik, Katarina, Elena Popova, Mihail Todiras, et al. "Inducible Transgenic Rat Model for Diabetes Mellitus Based on shRNA-Mediated Gene Knockdown." PLoS ONE 4, no. 4 (2009): e5124. http://dx.doi.org/10.1371/journal.pone.0005124.

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13

Azarnia, R., and W. R. Loewenstein. "Polyomavirus middle T antigen downregulates junctional cell-to-cell communication." Molecular and Cellular Biology 7, no. 2 (1987): 946–50. http://dx.doi.org/10.1128/mcb.7.2.946-950.1987.

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We examined the effect of polyomavirus middle T antigen on cell-to-cell communication in rat F cells transfected with an inducible middle T recombinant DNA or infected with a conditional mutant virus. Junctional permeability fell (reversibly) when middle T transcription was induced or when middle T was switched to the transformation+ form. The effect correlates with the rise of protein tyrosine kinase activity.
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14

Azarnia, R., and W. R. Loewenstein. "Polyomavirus middle T antigen downregulates junctional cell-to-cell communication." Molecular and Cellular Biology 7, no. 2 (1987): 946–50. http://dx.doi.org/10.1128/mcb.7.2.946.

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We examined the effect of polyomavirus middle T antigen on cell-to-cell communication in rat F cells transfected with an inducible middle T recombinant DNA or infected with a conditional mutant virus. Junctional permeability fell (reversibly) when middle T transcription was induced or when middle T was switched to the transformation+ form. The effect correlates with the rise of protein tyrosine kinase activity.
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15

Ouvrard-Pascaud, Antoine, Stefania Puttini, Yannis Sainte-Marie, et al. "Conditional gene expression in renal collecting duct epithelial cells: use of the inducible Cre-lox system." American Journal of Physiology-Renal Physiology 286, no. 1 (2004): F180—F187. http://dx.doi.org/10.1152/ajprenal.00301.2002.

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The renal collecting duct plays a key role in control of ion and fluid homeostasis. Genes encoding for ion transporters, hormone receptors, or regulatory proteins specifically expressed in the collecting duct are mutated in several genetic diseases with altered blood pressure. Suitable cellular models expressing genes in a conditional way should represent attractive systems for structure-function analyses and generation of appropriate physiopathological models of related diseases. However, generation of such systems remains laborious and quite inefficient. We adapted and improved a conditional
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16

Alonso, Lucille, Polina Peeva, Tania Fernández-del Valle Alquicira, et al. "Poor Decision Making and Sociability Impairment Following Central Serotonin Reduction in Inducible TPH2-Knockdown Rats." International Journal of Molecular Sciences 25, no. 9 (2024): 5003. http://dx.doi.org/10.3390/ijms25095003.

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Serotonin is an essential neuromodulator for mental health and animals’ socio-cognitive abilities. However, we previously found that a constitutive depletion of central serotonin did not impair rat cognitive abilities in stand-alone tests. Here, we investigated how a mild and acute decrease in brain serotonin would affect rats’ cognitive abilities. Using a novel rat model of inducible serotonin depletion via the genetic knockdown of tryptophan hydroxylase 2 (TPH2), we achieved a 20% decrease in serotonin levels in the hypothalamus after three weeks of non-invasive oral doxycycline administrati
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17

Lee, Sun Young, Ka Hyon Park, Gyuseok Lee та ін. "Hypoxia-inducible factor-2α mediates senescence-associated intrinsic mechanisms of age-related bone loss". Experimental & Molecular Medicine 53, № 4 (2021): 591–604. http://dx.doi.org/10.1038/s12276-021-00594-y.

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AbstractAging is associated with cellular senescence followed by bone loss leading to bone fragility in humans. However, the regulators associated with cellular senescence in aged bones need to be identified. Hypoxia-inducible factor (HIF)−2α regulates bone remodeling via the differentiation of osteoblasts and osteoclasts. Here, we report that HIF-2α expression was highly upregulated in aged bones. HIF-2α depletion in male mice reversed age-induced bone loss, as evidenced by an increase in the number of osteoblasts and a decrease in the number of osteoclasts. In an in vitro model of doxorubici
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18

Zhang, Huimin, Qi Zheng, and Ruby Yanru Chen-Tsai. "Establishment of a Cre-rat resource for creating conditional and physiological relevant models of human diseases." Transgenic Research 30, no. 1 (2021): 91–104. http://dx.doi.org/10.1007/s11248-020-00226-7.

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AbstractThe goal of this study is to establish a Cre/loxP rat resource for conditional and physiologically predictive rat models of human diseases. The laboratory rat (R. norvegicus) is a central experimental animal in several fields of biomedical research, such as cardiovascular diseases, aging, infectious diseases, autoimmunity, cancer models, transplantation biology, inflammation, cancer risk assessment, industrial toxicology, pharmacology, behavioral and addiction studies, and neurobiology. Up till recently, the ability of creating genetically modified rats has been limited compared to tha
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19

Hemmrich, Karsten, Christoph V. Suschek, Guido Lerzynski, Oliver Schnorr, and Victoria Kolb-Bachofen. "Specific iNOS-targeted antisense knockdown in endothelial cells." American Journal of Physiology-Cell Physiology 285, no. 2 (2003): C489—C498. http://dx.doi.org/10.1152/ajpcell.00045.2003.

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The inhibition of inducible nitric oxide synthase (iNOS) expression via antisense oligonucleotides (AS-ODN) may represent a highly specific tool. Endothelial cells (EC) represent prime candidate cells for in vivo application, and we therefore aimed at optimizing this technique for effectiveness and specificity in primary nontransformed rat EC. EC or L929 fibroblasts were incubated with iNOS-specific ODN optimizing all experimental steps. We find that ODN uptake, as analyzed by fluorescence microscopy and labeled ODN, was absolutely dependent on vehicle presence, and among the vehicles tested,
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20

Stahl, Andreas, Jean-Sebastian Joyal, Jing Chen, et al. "SOCS3 is an endogenous inhibitor of pathologic angiogenesis." Blood 120, no. 14 (2012): 2925–29. http://dx.doi.org/10.1182/blood-2012-04-422527.

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Abstract Inflammatory cytokines and growth factors drive angiogenesis independently; however, their integrated role in pathologic and physiologic angiogenesis is not fully understood. Suppressor of cytokine signaling-3 (SOCS3) is an inducible negative feedback regulator of inflammation and growth factor signaling. In the present study, we show that SOCS3 curbs pathologic angiogenesis. Using a Cre/Lox system, we deleted SOCS3 in vessels and studied developmental and pathologic angiogenesis in murine models of oxygen-induced retinopathy and cancer. Conditional loss of SOCS3 leads to increased pa
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21

De Conti, Giulia, Alicja M. Gruszka, Debora Valli, et al. "A Novel Platform to Test In Vivo Single Gene Dependencies in t(8,21) and t(15,17) AML Confirms Zeb2 as Leukemia Target." Cancers 12, no. 12 (2020): 3768. http://dx.doi.org/10.3390/cancers12123768.

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The increased usage of high-throughput technologies in cancer research, including genetic and drug screens, generates large sets of candidate targets that need to be functionally validated for their roles in tumor development. Thus, reliable and robust in vivo model systems are needed to perform reverse genetic experiments. Ideally, these models should allow for a conditional silencing of the target and an unambiguous identification of engineered cancer cells. Here, we present a platform consisting of: (i) t(8;21) and t(15;17) driven acute myeloid leukemia (AML) transgenic mice with constituti
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Zhao, Meng, Xu-Yun Zhao, Shi-Hua Liao, Ke-Wen Zhao, Li-Shun Wang та Guo-Qiang Chen. "The Potential Role of Galectin-1 in Hypoxia Induced AML Cell Differentiation through C/EBPαa Transcription Regulation." Blood 114, № 22 (2009): 3120. http://dx.doi.org/10.1182/blood.v114.22.3120.3120.

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Abstract Abstract 3120 Poster Board III-57 Recently we reported that cobalt chloride-simulated hypoxia and mild hypoxia could induce the differentiation of human acute myeloid leukemic (AML) cells probably through hypoxia inducible factor-1 alpha (HIF-1αa) independent of its transcription activity. In this study, we used differential gel electrophoresis to compare the HIF-1αa regulated proteins between conditional induction and absence of HIF-1αa protein in U937T cells. Among all the up-regulated proteins indentified, the mRNA and protein level of galectin-1 was most remarkably changed. Knockd
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23

Reichhart, Nadine, Sergio Crespo-Garcia, Nadine Haase, et al. "The TetO rat as a new translational model for type 2 diabetic retinopathy by inducible insulin receptor knockdown." Diabetologia 60, no. 1 (2016): 202–11. http://dx.doi.org/10.1007/s00125-016-4115-0.

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24

Kanyo, R., D. M. Price, C. L. Chik, and A. K. Ho. "Salt-Inducible Kinase 1 in the Rat Pinealocyte: Adrenergic Regulation and Role in Arylalkylamine N-Acetyltransferase Gene Transcription." Endocrinology 150, no. 9 (2009): 4221–30. http://dx.doi.org/10.1210/en.2009-0275.

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Abstract The recognition of the basic leucine zipper domain in the regulation of transcriptional activity of cAMP response element-binding protein by salt-inducible kinase (SIK) prompted our investigation of the regulatory role of this kinase in the induction of Aa-nat and other cAMP-regulated genes in the rat pineal gland. Here we report Sik1 expression was induced by norepinephrine (NE) in rat pinealocytes primarily through activation of β-adrenergic receptors, with a minor contribution from activation of α-adrenergic receptors. Treatments with dibutyryl cAMP, and to a lesser extent, agents
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25

Ameis, Dustin, Franklin Liu, Eimear Kirby, Daywin Patel, and Richard Keijzer. "The RNA-binding protein Quaking regulates multiciliated and basal cell abundance in the developing lung." American Journal of Physiology-Lung Cellular and Molecular Physiology 320, no. 4 (2021): L557—L567. http://dx.doi.org/10.1152/ajplung.00481.2019.

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RNA-binding proteins (RBPs) form complexes with RNA, changing how the RNA is processed and thereby regulating gene expression. RBPs are important sources of gene regulation during organogenesis, including the development of lungs. The RBP called Quaking (QK) is critical for embryogenesis, yet it has not been studied in the developing lung. Here, we show that QK is widely expressed during rat lung development and into adulthood. The QK isoforms QK5 and QK7 colocalize to the nuclei of nearly all lung cells. QK6 is present in the nuclei and cytoplasm of mesenchymal cells and is only present in th
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Zhan, Wenxing, Liping Chen, Hongfei Liu, et al. "Pcsk6 Deficiency Promotes Cardiomyocyte Senescence by Modulating Ddit3-Mediated ER Stress." Genes 13, no. 4 (2022): 711. http://dx.doi.org/10.3390/genes13040711.

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Cardiac aging is a critical determinant of cardiac dysfunction, which contributes to cardiovascular disease in the elderly. Proprotein convertase subtilisin/kexin 6 (PCSK6) is a proteolytic enzyme important for the maintenance of cardiac function and vascular homeostasis. To date, the involvement of PCSK6 in cardiac aging remains unknown. Here we report that PCSK6 expression decreased in the hearts of aged mice, where high levels cyclin dependent kinase inhibitor 2A (P16) and cyclin dependent kinase inhibitor 1A (P21) (senescence markers) were observed. Moreover, PCSK6 protein expression was s
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Fang, Yi, Xiaofang Yu, Yong Liu та ін. "miR-29c is downregulated in renal interstitial fibrosis in humans and rats and restored by HIF-α activation". American Journal of Physiology-Renal Physiology 304, № 10 (2013): F1274—F1282. http://dx.doi.org/10.1152/ajprenal.00287.2012.

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Treatment with l-mimosine, which activates hypoxia-inducible factor-α (HIF-α), attenuates renal tubulointerstitial injury and improves renal function in a rat remnant kidney model. The miR-29 family of microRNAs directly targets a large number of extracellular matrix genes and reduces renal interstitial fibrosis. We analyzed microRNA expression profiles in rat remnant kidneys with or without treatment with l-mimosine. The expression of miR-29c was downregulated in rat remnant kidneys compared with sham control and significantly restored by the l-mimosine treatment. In cultured human kidney epi
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28

Giles, Jasmine, Jitandrakumar R. Patel, Adam Miller, Elizabeth Iverson, Daniel Fitzsimons, and Richard L. Moss. "Recovery of left ventricular function following in vivo reexpression of cardiac myosin binding protein C." Journal of General Physiology 151, no. 1 (2018): 77–89. http://dx.doi.org/10.1085/jgp.201812238.

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The loss of cardiac myosin binding protein C (cMyBP-C) results in left ventricular dilation, cardiac hypertrophy, and impaired ventricular function in both constitutive and conditional cMyBP-C knockout (MYBPC3 null) mice. It remains unclear whether the structural and functional phenotypes expressed in the MYBPC3 null mouse are reversible, which is an important question, since reduced expression of cMyBP-C is an important cause of hypertrophic cardiomyopathy in humans. To investigate this question, we generated a cardiac-specific transgenic mouse model using a Tet-Off inducible system to permit
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Chu, Miensheng, Yevgeniya Koshman, Rekha Iyengar, Taehoon Kim, Brenda Russell, and Allen M. Samarel. "Contractile Activity Regulates Inducible Nitric Oxide Synthase Expression and NOi Production in Cardiomyocytes via a FAK-Dependent Signaling Pathway." Journal of Signal Transduction 2012 (July 26, 2012): 1–11. http://dx.doi.org/10.1155/2012/473410.

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Intracellular nitric oxide (NOi) is a physiological regulator of excitation-contraction coupling, but is also involved in the development of cardiac dysfunction during hypertrophy and heart failure. To determine whether contractile activity regulates nitric oxide synthase (NOS) expression, spontaneously contracting, neonatal rat ventricular myocytes (NRVM) were treat with L-type calcium channel blockers (nifedipine and verapamil) or myosin II ATPase inhibitors (butanedione monoxime (BDM) and blebbistatin) to produce contractile arrest. Both types of inhibitors significantly reduced iNOS but no
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30

Azevedo, Mauro F., Paul R. Sanders, Efrosinia Krejany, et al. "Inhibition of Plasmodium falciparum CDPK1 by conditional expression of its J-domain demonstrates a key role in schizont development." Biochemical Journal 452, no. 3 (2013): 433–41. http://dx.doi.org/10.1042/bj20130124.

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PfCDPK1 [Plasmodium falciparum CDPK1 (calcium-dependent protein kinase 1)] is highly expressed in parasite asexual blood and mosquito stages. Its role is still poorly understood, but unsuccessful gene knockout attempts suggest that it is essential for parasite replication and/or RBC (red blood cell) invasion. In the present study, by tagging endogenous CDPK1 with GFP (green fluorescent protein), we demonstrate that CDPK1 localizes to the parasite plasma membrane of replicating and invasive forms as well as very young intracellular parasites and does not appear to be exported into RBCs. Althoug
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31

Lin, En-Shyh, Yu-An Hsu, Ching-Yao Chang, Hui-Ju Lin, Chih Sheng Chen, and Lei Wan. "Ablation of Galectin-12 Inhibits Atherosclerosis through Enhancement of M2 Macrophage Polarization." International Journal of Molecular Sciences 21, no. 15 (2020): 5511. http://dx.doi.org/10.3390/ijms21155511.

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The formation of foam cells, which are macrophages that have engulfed oxidized low-density lipoprotein (OxLDL), constitutes the first stage in the development of atherosclerosis. Previously, we found that knocking down galectin-12, a negative regulator of lipolysis, leads to reduced secretion of monocyte chemoattractant protein-1 (MCP-1), a chemokine that plays an important role in atherosclerosis. This prompted us to study the role of galectin-12 in atherosclerosis. With that aim, we examined foam cell formation in Gal12‒/‒ murine macrophages exposed to OxLDL and acetylated LDL (AcLDL). Then,
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Herr, Barbara, Jie Zhou, Christian Werno та ін. "The supernatant of apoptotic cells causes transcriptional activation of hypoxia-inducible factor–1α in macrophages via sphingosine-1-phosphate and transforming growth factor-β". Blood 114, № 10 (2009): 2140–48. http://dx.doi.org/10.1182/blood-2009-01-201889.

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Abstract Macrophages infiltrating solid tumors exhibit a tumor-supporting phenotype and are critical for tumor development. Little is known which tumor-derived signal provokes this phenotype shift and how these signals are interpreted in macrophages to support tumor growth. We used the supernatant of apoptotic cells and noticed transcriptional, nuclear factor of activated T cells-dependent up-regulation of hypoxia-inducible factor (HIF)–1α mRNA, subsequent protein expression, and HIF-1 activity. Blocking calcineurin with cyclosporine A attenuated nuclear factor of activated T cells binding dur
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Yan, Xiaoying, Ran Zhao, Xiaorong Feng та ін. "Sialyltransferase7A promotes angiotensin II-induced cardiomyocyte hypertrophy via HIF-1α-TAK1 signalling pathway". Cardiovascular Research 116, № 1 (2019): 114–26. http://dx.doi.org/10.1093/cvr/cvz064.

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Abstract Aims Sialylation is up-regulated during the development of cardiac hypertrophy. Sialyltransferase7A (Siat7A) mRNA is consistently over-expressed in the hypertrophic left ventricle of hypertensive rats independently of genetic background. The aims of this study were: (i) to detect the Siat7A protein levels and its roles in the pathological cardiomyocyte hypertrophy; (ii) to elucidate the effect of sialylation mediated by Siat7A on the transforming-growth-factor-β-activated kinase (TAK1) expression and activity in cardiomyocyte hypertrophy; and (iii) to clarify hypoxia-inducible factor
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Shibahara, Daisuke Jefferson, Naoki Akanuma, Eunyoung Heo, et al. "Abstract 3723: Role of KAT5 in non-small cell lung cancer." Cancer Research 82, no. 12_Supplement (2022): 3723. http://dx.doi.org/10.1158/1538-7445.am2022-3723.

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Abstract Histone modifications play crucial roles in transcriptional activation, and aberrant epigenetic changes are associated with oncogenesis. Lysine (K) acetyltransferases 5 (KAT5) is reportedly implicated in cancer development and maintenance, although the role of KAT5 in lung cancer largely remain unclear. Here we demonstrate that KAT5 expression was lower in lung adenocarcinoma than normal lung cells. Overexpression of KAT5 had no effect on tumor proliferation and migration, whereas knockdown of KAT5 paradoxically decreased those activities. To assess KAT5 function in lung tumorigenesis
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Figueiredo, Marta, Arezoo Daryadel, Gabin Sihn, et al. "The (pro)renin receptor (ATP6ap2) facilitates receptor-mediated endocytosis and lysosomal function in the renal proximal tubule." Pflügers Archiv - European Journal of Physiology 473, no. 8 (2021): 1229–46. http://dx.doi.org/10.1007/s00424-021-02598-z.

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AbstractThe ATP6ap2 (Pro)renin receptor protein associates with H+-ATPases which regulate organellar, cellular, and systemic acid–base homeostasis. In the kidney, ATP6ap2 colocalizes with H+-ATPases in various cell types including the cells of the proximal tubule. There, H+-ATPases are involved in receptor-mediated endocytosis of low molecular weight proteins via the megalin/cubilin receptors. To study ATP6ap2 function in the proximal tubule, we used an inducible shRNA Atp6ap2 knockdown rat model (Kd) and an inducible kidney-specific Atp6ap2 knockout mouse model. Both animal lines showed highe
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36

Csiszar, Anna, Nazar Labinskyy, Andrej Podlutsky, et al. "Vasoprotective effects of resveratrol and SIRT1: attenuation of cigarette smoke-induced oxidative stress and proinflammatory phenotypic alterations." American Journal of Physiology-Heart and Circulatory Physiology 294, no. 6 (2008): H2721—H2735. http://dx.doi.org/10.1152/ajpheart.00235.2008.

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The dietary polyphenolic compound resveratrol, by activating the protein deacetylase enzyme silent information regulator 2/sirtuin 1 (SIRT1), prolongs life span in evolutionarily distant organisms and may mimic the cytoprotective effects of dietary restriction. The present study was designed to elucidate the effects of resveratrol on cigarette smoke-induced vascular oxidative stress and inflammation, which is a clinically highly relevant model of accelerated vascular aging. Cigarette smoke exposure of rats impaired the acetylcholine-induced relaxation of carotid arteries, which could be preven
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Abraham, Ajay, Puneet Agarwal, Hui Li, Andrew Paterson, Jianbo He, and Ravi Bhatia. "Inhibition of CML Development Following Conditional SIRT1 Deletion in Transgenic BCR-ABL Mice." Blood 128, no. 22 (2016): 931. http://dx.doi.org/10.1182/blood.v128.22.931.931.

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Abstract Despite the success of tyrosine kinase inhibitors (TKIs) in treatment of CML, cures remain elusive, as primitive leukemia stem cells (LSC) are retained in patients achieving remission. Previous studies from our group have suggested that Sirtuin 1 (SIRT1) inhibition may represent a novel approach for elimination of LSCs in chronic phase CML. SIRT1 was shown to be overexpressed in CML LSCs, and SIRT1 inhibition using shRNA or a small molecule SIRT1 inhibitor selectively eliminated CML LSCs by increasing p53 acetylation and activity (Li et.al; Cancer Cell 2012). These studies were limite
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38

Vukovic, Milica, Amelie V. Guitart, Catarina Sepulveda та ін. "Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance". Journal of Experimental Medicine 212, № 13 (2015): 2223–34. http://dx.doi.org/10.1084/jem.20150452.

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Leukemogenesis occurs under hypoxic conditions within the bone marrow (BM). Knockdown of key mediators of cellular responses to hypoxia with shRNA, namely hypoxia-inducible factor-1α (HIF-1α) or HIF-2α, in human acute myeloid leukemia (AML) samples results in their apoptosis and inability to engraft, implicating HIF-1α or HIF-2α as therapeutic targets. However, genetic deletion of Hif-1α has no effect on mouse AML maintenance and may accelerate disease development. Here, we report the impact of conditional genetic deletion of Hif-2α or both Hif-1α and Hif-2α at different stages of leukemogenes
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39

Duque, Cristiane, Rafael N. Stipp, Bing Wang, et al. "Downregulation of GbpB, a Component of the VicRK Regulon, Affects Biofilm Formation and Cell Surface Characteristics ofStreptococcus mutans." Infection and Immunity 79, no. 2 (2010): 786–96. http://dx.doi.org/10.1128/iai.00725-10.

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ABSTRACTThe virulence of the dental caries pathogenStreptococcus mutansrelies in part on the sucrose-dependent synthesis of and interaction with glucan, a major component of the extracellular matrix of tooth biofilms. However, the mechanisms by which secreted and/or cell-associated glucan-binding proteins (Gbps) produced byS. mutansparticipate in biofilm growth remain to be elucidated. In this study, we further investigate GbpB, an essential immunodominant protein with similarity to murein hydrolases. A conditional knockdown mutant that expressedgbpBantisense RNA under the control of a tetracy
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Brown, Stephen T., та Colin A. Nurse. "Induction of HIF-2α is dependent on mitochondrial O2 consumption in an O2-sensitive adrenomedullary chromaffin cell line". American Journal of Physiology-Cell Physiology 294, № 6 (2008): C1305—C1312. http://dx.doi.org/10.1152/ajpcell.00007.2008.

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During low O2 (hypoxia), hypoxia-inducible factor (HIF)-α is stabilized and translocates to the nucleus, where it regulates genes critical for survival and/or adaptation in low O2. While it appears that mitochondria play a critical role in HIF induction, controversy surrounds the underlying mechanism(s). To address this, we monitored HIF-2α expression and oxygen consumption in an O2-sensitive immortalized rat adrenomedullary chromaffin (MAH) cell line. Hypoxia (2–8% O2) caused a concentration- and time-dependent increase in HIF-2α induction, which was blocked in MAH cells with either RNA inter
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41

Weaver, Ian C. G., Ian C. Hellstrom, Shelley E. Brown, et al. "The methylated-DNA binding protein MBD2 enhances NGFI-A (egr-1)-mediated transcriptional activation of the glucocorticoid receptor." Philosophical Transactions of the Royal Society B: Biological Sciences 369, no. 1652 (2014): 20130513. http://dx.doi.org/10.1098/rstb.2013.0513.

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Variations in maternal care in the rat influence the epigenetic state and transcriptional activity of glucocorticoid receptor (GR) gene in the hippocampus. The mechanisms underlying this maternal effect remained to be defined, including the nature of the relevant maternally regulated intracellular signalling pathways. We show here that increased maternal licking/grooming (LG), which stably enhances hippocampal GR expression, paradoxically increases hippocampal expression of the methyl-CpG binding domain protein-2 (MBD2) and MBD2 binding to the exon 1 7 GR promoter. Knockdown experiments of MBD
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42

Chakraborty, Damayanti, Wei Cui, Gracy X. Rosario, et al. "HIF-KDM3A-MMP12 regulatory circuit ensures trophoblast plasticity and placental adaptations to hypoxia." Proceedings of the National Academy of Sciences 113, no. 46 (2016): E7212—E7221. http://dx.doi.org/10.1073/pnas.1612626113.

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The hemochorial placenta develops from the coordinated multilineage differentiation of trophoblast stem (TS) cells. An invasive trophoblast cell lineage remodels uterine spiral arteries, facilitating nutrient flow, failure of which is associated with pathological conditions such as preeclampsia, intrauterine growth restriction, and preterm birth. Hypoxia plays an instructive role in influencing trophoblast cell differentiation and regulating placental organization. Key downstream hypoxia-activated events were delineated using rat TS cells and tested in vivo, using trophoblast-specific lentivir
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Kobayashi, Eri, Ritsuko Shimizu, Yuko Kikuchi, Satoru Takahashi, and Masayuki Yamamoto. "Loss of the Gata1 Gene IE Exon Leads to Variant Transcript Expression and the Production of GATA1 Protein Lacking the N-Terminal Domain." Blood 114, no. 22 (2009): 3648. http://dx.doi.org/10.1182/blood.v114.22.3648.3648.

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Abstract Abstract 3648 Poster Board III-584 GATA1 is a transcription factor essential for the differentiation of erythroid cells and megakaryocytes. Since GATA1 regulates genes related to the survival, proliferation and differentiation of hematopoietic cells, regulation of the Gata1 gene expression is critically important for the understanding of hematopoiesis. The Gata1 locus contains multiple untranslated first exons plus five common coding exons. Of these first exons, erythroid first exon (IE exon) is important for the Gata1 gene expression in the hematopoietic lineages. However, due to the
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44

McTague, J., M. Ferguson, C. L. Chik, and A. K. Ho. "Sustained Adrenergic Stimulation Is Required for the Nuclear Retention of TORC1 in Male Rat Pinealocytes." Endocrinology 154, no. 9 (2013): 3240–50. http://dx.doi.org/10.1210/en.2013-1293.

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The process involved in relocation of the coactivator, transducer of regulated cAMP-regulated element-binding protein (TORC) to the cytoplasm, unlike its activation, is not well understood. Using cultured pineal cells prepared from male rats, we found that although both α- and β-adrenergic stimulation could cause TORC1 dephosphorylation, only α-adrenergic stimulation was effective in the norepinephrine (NE)-mediated translocation of TORC1 into the nucleus. In contrast, blockade of either the α- or the β-adrenergic receptor after NE stimulation was effective in causing the rephosphorylation and
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Zhang, Wei, Xiangjun Zhou, Qisheng Yao, Yutao Liu, Hao Zhang, and Zheng Dong. "HIF-1-mediated production of exosomes during hypoxia is protective in renal tubular cells." American Journal of Physiology-Renal Physiology 313, no. 4 (2017): F906—F913. http://dx.doi.org/10.1152/ajprenal.00178.2017.

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Exosomes are nano-sized vesicles produced and secreted by cells to mediate intercellular communication. The production and function of exosomes in kidney tissues and cells remain largely unclear. Hypoxia is a common pathophysiological condition in kidneys. This study was designed to characterize exosome production during hypoxia of rat renal proximal tubular cells (RPTCs), investigate the regulation by hypoxia-inducible factor-1 (HIF-1), and determine the effect of the exosomes on ATP-depletion-induced tubular cell injury. Hypoxia did not change the average sizes of exosomes secreted by RPTCs,
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46

Wang, Wei, Andrew P. Landstrom, Qiongling Wang, et al. "Reduced junctional Na+/Ca2+-exchanger activity contributes to sarcoplasmic reticulum Ca2+ leak in junctophilin-2-deficient mice." American Journal of Physiology-Heart and Circulatory Physiology 307, no. 9 (2014): H1317—H1326. http://dx.doi.org/10.1152/ajpheart.00413.2014.

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Expression silencing of junctophilin-2 (JPH2) in mouse heart leads to ryanodine receptor type 2 (RyR2)-mediated sarcoplasmic reticulum (SR) Ca2+ leak and rapid development of heart failure. The mechanism and physiological significance of JPH2 in regulating RyR2-mediated SR Ca2+ leak remains elusive. We sought to elucidate the role of JPH2 in regulating RyR2-mediated SR Ca2+ release in the setting of cardiac failure. Cardiac myocytes isolated from tamoxifen-inducible conditional knockdown mice of JPH2 (MCM-shJPH2) were subjected to confocal Ca2+ imaging. MCM-shJPH2 cardiomyocytes exhibited an i
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47

Resnitzky, D., M. Gossen, H. Bujard, and S. I. Reed. "Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system." Molecular and Cellular Biology 14, no. 3 (1994): 1669–79. http://dx.doi.org/10.1128/mcb.14.3.1669-1679.1994.

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Conditional overexpression of human cyclins B1, D1, and E was accomplished by using a synthetic cDNA expression system based on the Escherichia coli tetracycline repressor. After induction of these cyclins in asynchronous Rat-1 fibroblasts, a decrease in the length of the G1 interval was observed for cyclins D1 and E, consistent with an acceleration of the G1/S phase transition. We observed, in addition, a compensatory lengthening of S phase and G2 so that the mean cell cycle length in populations constitutively expressing these cyclins was unchanged relative to those of their uninduced counte
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48

Resnitzky, D., M. Gossen, H. Bujard, and S. I. Reed. "Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system." Molecular and Cellular Biology 14, no. 3 (1994): 1669–79. http://dx.doi.org/10.1128/mcb.14.3.1669.

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Conditional overexpression of human cyclins B1, D1, and E was accomplished by using a synthetic cDNA expression system based on the Escherichia coli tetracycline repressor. After induction of these cyclins in asynchronous Rat-1 fibroblasts, a decrease in the length of the G1 interval was observed for cyclins D1 and E, consistent with an acceleration of the G1/S phase transition. We observed, in addition, a compensatory lengthening of S phase and G2 so that the mean cell cycle length in populations constitutively expressing these cyclins was unchanged relative to those of their uninduced counte
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49

Ren, Kehan, Zongjun Xia, Ermin Li, Xu Han, and Peng Ji. "Rapid Degradation of mDia2 Protein during Terminal Erythropoiesis Via an In Vivo Aid System: An Alternative Approach for Loss-of-Function Studies." Blood 142, Supplement 1 (2023): 2443. http://dx.doi.org/10.1182/blood-2023-178268.

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Loss-of-function manipulations are crucial methods for studying gene functions. Despite the availability of many chemical-induced genetic manipulation techniques, there are limitations associated with these technologies. Tamoxifen-induced conditional gene expression, gene knockdown, or cell tracking represent some of the most common genetic manipulations employed in mice. However, tamoxifen-induced gene or Cre expression in bone marrow cell populations is relatively weak, affecting only a low proportion of cells. Similar observations have been made with the in vivo doxycycline-inducible system
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Querbes, William, Roman Bogorad, Javid Moslehi, et al. "Liver Specific Delivery of siRNA Targeting EGLN Prolyl Hydroxylases Activates Hepatic Erythropoietin Production and Stimulates Erythropoiesis,." Blood 118, no. 21 (2011): 3161. http://dx.doi.org/10.1182/blood.v118.21.3161.3161.

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Abstract Abstract 3161 Anemia in chronic kidney disease patients due to impaired renal production of erythropoietin (EPO) and insufficient erythropoiesis is a significant public health problem. One approach to compensate for the low EPO levels in patients with impaired kidney function would be to stimulate the levels of EPO production from non-renal sources. It is well known that during fetal development the liver serves as the primary producer of EPO until the kidney becomes the dominant source after birth. Negative regulation of EPO production is mediated by the EGLN family of prolyl hydroxy
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