Research Paper Volume 13, Issue 9 pp 12780—12799

CircRNA_30032 promotes renal fibrosis in UUO model mice via miRNA-96-5p/HBEGF/KRAS axis

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Figure 5. CircRNA_30032 directly binds to miR-96-5p in BUMPT cells and murine kidney tissues. (A) RegRNA 2.0 software analysis shows putative miR-96-5p binding site in the circRNA_30032 sequence. (B) Dual luciferase reporter assay results show relative luciferase activities in BUMPT cells co-transfected with circRNA_30032-WT (wild-type) or circRNA_30032-MT (mutant) plus miR-96-5p or scrambled miRNA. (C, D) RNA FISH images show co-localization of circRNA_30032 and miR-96-5p in kidney tissue sections from UUO-induced C57BL/6 mice and TGF-β1-treated BUMPT cells. (E, F) RT-qPCR analysis shows miR-96-5p expression levels in control, circRNA_30032-silenced, and circRNA_30032-overexpressing BUMPT cells treated with or without TGF-β1 for 24 h. Note: # denotes p<0.05 when comparing scrambled miRNA plus TGF-β1 or circRNA_30032 plus saline groups vs. scrambled miRNA plus saline group; * denotes p<0.05 when comparing circRNA_30032 WT plus miR-96-5p mimic vs. other groups, circRNA_30032 siRNA plus TGF-β1 group vs. scrambled miRNA plus TGF-β1 group.