Fig. 6

Increased WTAP expression promotes the N6-methylation and stabilization of ASC mRNA in GCs from PCOS mice. A Methylated RNA Immunoprecipitation (Me-RIP) assay to quantify m6A level of ASC mRNA in GCs from control, PCOS and Plumbagin-treated mice. N = 4. B Relative expression of m6A-related genes in GCs from three experimental mice groups. n = 4. C WTAP transcripts in the GCs of control and PCOS mice were quantified by RT-PCR. D WTAP protein in control and GCs were detected by immunoblot. E Relative WTAP protein level normalized with β-actin. N = 3. F Overexpress WTAP in GC cell line HGL5 by transfecting with a recombinant plasmid containing WTAP. The relative expression of ASC mRNA in HGL5 was determined by RT-PCR. n = 4. G Relative m6A level of ASC mRNA level in HGL5 cells were enriched and quantified by Me-RIP. n = 4. Individual data points are displayed. Data are mean ± SEM. one-way ANOVA were used to analysis the difference in A,F,G; student t-test was used to analysis difference in C and E. *P < 0.05; **P < 0.01; ***P < 0.001