Figure 5.

CircYTHDC2 negatively regulates TET2. (A) The schematic diagram of TET2-WT or TET2-Mut luciferase reporter gene. The wild type motifs were indicated by green and the mutant motif was indicated by red. A7R5 cells were transfected with TET2-WT or TET2-Mut, combined with circYTHDC2 siRNA. The cells transfected with scramble control were used as control. Luciferase report gene assay were performed to measure the relative luciferase activity. (B) Relative enrichment representing TET2 and circYTHDC2 RNA levels associated with circYTHDC2 junction compared to control. (C) qRT–PCR analysis for the expression of TET2 after high glucose treatment in A7R5 cells. (D) western blotting analysis of TET2 expression after high glucose treatment in A7R5 cells. (E) qRT–PCR analysis for the expression of TET2 after circYTHDC2 siRNA transfection in A7R5 cells. (F) western blotting analysis of TET2 expression after circYTHDC2 siRNA transfection in A7R5 cells. (G) western blotting analysis of TET2 expression after TET2 siRNA transfection in A7R5 cells. (H) western blotting analysis for the expression of MYODC, SRF and KLF4 in A7R5 cells after circYTHDC2 siRNA transfection alone, or combined with TET2 siRNA transfection. (I) qRT–PCR analysis for the expression of TET2 after circYTHDC2 transfection in A7R5 cells. (J) western blotting analysis of TET2 expression after circYTHDC2 transfection in A7R5 cells. (K) western blotting analysis of TET2 expression after TET2 transfection in A7R5 cells. (L) western blotting analysis for the expression of MYODC, SRF and KLF4 in A7R5 cells after circYTHDC2 transfection alone, or combined with TET2 transfection. Three independent studies were performed and the data were expressed as mean ± standard deviation. HG, high glucose. *p < 0.05, **p < 0.01.