Fig. 7
From: Circular RNA-circPan3 attenuates cardiac hypertrophy via miR-320-3p/HSP20 axis
ALKBH5 regulates the stability of circPan3 through m6A modification. A The level of N6-methylated circPan3 in cardiomyocytes is increased by ISO-stimulation. The lysate of cardiomyocytes treated with ISO or saline (con) was incubated with m6A antibody or control IgG-coated agarose beads. The N6-methylated RNAs captured by antibodies were purified by TRIzol. The circPan3 level was analyzed by qPCR. *P < 0.05. n = 3. B, C ALKBH5 is significantly downregulated in cardiomyocyte hypertrophy. Cardiomyocytes were treated with 10 μΜ ISO for 24 h. The protein and mRNA levels of m6A writers and erasers in cardiomyocytes were analyzed by B immunoblotting and C qPCR, respectively. **P < 0.01. n = 3. D–E Knockdown of ALKBH5 reduces the expression of circPan3 in cardiomyocytes. Cardiomyocytes were transfected with ALKBH5-siRNA for 24 h. The expression of ALKBH5 D and circPan3 E was measured by qPCR. *P < 0.05, **P < 0.01. n = 3. F–I Silencing of ALKBH5 promotes cardiomyocyte hypertrophy. Cardiomyocyte hypertrophy was assessed by F cell surface area measurement and level of G ANP, H BNP and I β-MHC. The expression level of ANP, BNP and β-MHC was analyzed by qPCR. *P < 0.05, **P < 0.01. n = 3. J–N Overexpression of circPan3 abolishes cardiomyocyte hypertrophy induced by knockdown of ALKBH5. J Cardiomyocytes were co-transfected with ALKBH5-siRNA and the circPan3 vector for 24 h. The expression of circPan3 was measured by qPCR. *P < 0.05, **P < 0.01. n = 3. Cardiomyocyte hypertrophy was assessed by K cell surface area measurement and level of L ANP, M BNP and N β-MHC. The expression level of ANP, BNP and β-MHC was analyzed by qPCR. *P < 0.05, **P < 0.01. n = 3