Fig. 5
circMAST1 binds with NAT10 and regulates ac4C modification of YAP mRNA. A Mass spectrometry identified NAT10 was pulled down by circMAST1 from SiHa cells lysates. B, C The interaction between circMAST1 and NAT10 in SiHa and HeLa cells was shown by pull-down assay (B) and RIP assay (C). D The predicted secondary structure of circMAST1 using the RNAfold WebServer, based on the minimum free energy. Color scales indicated the confidence of predictions for each base, and the red shades demonstrated the predictions with strong confidence. E RNA pulldown assay showed NAT10 pulled down by biotin-labeled circMAST1 of different lengths. F, G Prediction of conserved acetylation site on YAP mRNA by PACES tool. H Relative protein expression levels of YAP were analyzed by western blotting after NAT10 overexpression or knockdown. I, J The regulatory role of circMAST1 on YAP ac4C in SiHa and HeLa cells confirmed by the acRIP-qPCR assay in the indicated groups. K, L The impact of circMAST1 on YAP mRNA stability confirmed by the RNA decay assay. Each experiment was performed at least three times independently. **P < 0.01. *** P < 0.001. ****P < 0.0001