Fig. 6: ALKBH5 regulates STAT3 activity in a m6A-dependent manner.
From: The functional role of m6A demethylase ALKBH5 in cardiomyocyte hypertrophy
A Relative enrichment of Stat3 mRNA associated with ALKBH5 protein was identified by RIP-qPCR assays using anti-ALKBH5 antibody. n = 4. B MeRIP-qPCR assays were performed to detect the relative m6A enrichment of Stat3 mRNA in PE-induced H9C2 cells. n = 4. C The online prediction of Stat3 mRNA in SRAMP website showed the potential sites of m6A modification. The purple boxes represent m6A sites with high confidence. D RIP-qPCR analysis of the enrichment of ALKBH5 protein on predicted m6A modification sites of Stat3 mRNA CDS and 3’-UTR regions (Stat3 CDS-1318, Stat3 3’-UTR-2953, Stat3 3’-UTR-3153) in H9C2 cells. E m6A modification of Stat3 3’-UTR-2953 was detected by MeRIP-qPCR analysis using m6A antibody in Alkbh5 OE-transfected H9C2 cells upon PE treatment. F The mutation at the putative m6A site (2953) in Stat3 3’-UTR (A to G) was generated and the Stat3 mRNA coding sequence (pGL3-WT) and Stat3 3’-UTR-2953 mutated sequence (pGL3-Mut) luciferase reporter vectors were constructed. G Relative luciferase activity of the pGL3-empty, pGL3-WT, and pGL-Mut luciferase reporter in Control and Alkbh5 OE-induced H9C2 cells. Data are expressed as the mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.
