%0 Figure %A N., Kushida %A S., Nomura %A I., Mimura %A T., Fujita %A S., Yamamoto %A M., Nangaku %A H., Aburatani %D 2016 %T PowerPoint Slides for: Hypoxia-Inducible Factor-1α Activates the Transforming Growth Factor-β/SMAD3 Pathway in Kidney Tubular Epithelial Cells %U https://karger.figshare.com/articles/figure/PowerPoint_Slides_for_Hypoxia-Inducible_Factor-1_Activates_the_Transforming_Growth_Factor-_SMAD3_Pathway_in_Kidney_Tubular_Epithelial_Cells/3823917 %R 10.6084/m9.figshare.3823917.v1 %2 https://karger.figshare.com/ndownloader/files/5961177 %K Chromatin immunoprecipitation-sequencing %K Hypoxia-inducible factor-1α %K Hypoxia %K Kidney tubular epithelial cell %K RNA-sequencing %K SMAD3 %K Transforming growth factor-β %X Background: Kidney injury, including chronic kidney disease and acute kidney injury, is a worldwide health problem. Hypoxia and transforming growth factor-β (TGF-β) are well-known factors that promote kidney injury. Hypoxia-inducible factor (HIF) and SMAD3 are their main downstream transcriptional factors. Hypoxia-HIF pathway and TGF-β/SMAD3 pathway play a crucial role in the progression of kidney injury. However, reports on their interactions are limited, and the global transcriptional regulation under their control is almost unknown. Methods: Kidney tubular epithelial cells were cultured and stimulated by hypoxia and TGF-β. We detected global binding sites of HIF-1α and SMAD3 in cells using chromatin immunoprecipitation-sequencing (ChIP-Seq), and measured the gene expression using RNA-sequencing (RNA-Seq). ChIP-quantitative PCR (qPCR) was used to quantitatively evaluate bindings of SMAD3. Results: ChIP-Seq revealed that 2,065 and 5,003 sites were bound by HIF-1α and SMAD3, respectively, with 614 sites co-occupied by both factors. RNA-Seq showed that hypoxia and TGF-β stimulation causes synergistic upregulation of 249 genes, including collagen type I alpha 1 (COL1A1) and serpin peptidase inhibitor, clade E, member 1, which are well-known to be involved in fibrosis. Ontology of the 249 genes implied that the interaction of HIF-1α and SMAD3 is related to biological processes such as fibrosis. ChIP-qPCR of SMAD3 at HIF-1α binding sites near COL1A1 and SERPINE1 indicated that HIF-1α promotes the bindings of SMAD3, which is induced by TGF-β. Conclusions: These findings suggest that HIF-1α induced by hypoxia activates the TGF-β/SMAD3 pathway. This mechanism may promote kidney injury, especially by upregulating genes related to fibrosis. %I Karger Publishers