Fig. 1
Ac4C regulates mRNA stability and translation of YTHDC1. A U2OS and 143B cells were transfected with negative control (NC) siRNA or NAT10 siRNA for 24 h. The knockdown efficiency of NAT10 siRNA in U2OS and 143B cells was evaluated by qRT-PCR analysis. B The specificity of the ac4C antibody was detected in 143B cells by Dot blot analyses. C The m6A content in 143B cells was detected by ELISA assay. D Protein expression of NAT10, METTL3, METTL14, ALKBH5, YTHDF1, YTHDF2, IGF2BP1, IGF2BP2, IGF2BP3, YTHDC1 in 143B cells was detected by western blotting. E Prediction diagram of YTHDC1 mRNA acetylation site. F qRT-PCR analysis revealed the expression of YTHDC1 after knockdown of NAT10 in U2OS and 143B cells. G RIP-qPCR assay was performed to analyze the mRNA levels of ac4C-modified in 143B cells with or without knockdown of NAT10. H Luciferase activity assay was performed to confirm the YTHDC1 mRNA was directly bound to NAT10 in HEK-293 T cells. I The YTHDC1 mRNA half-life was estimated according to linear regression analysis after the indicated actinomycin D treatment. J 143B cells were transfected with negative control (NC) siRNA or NAT10 siRNA for 24 h and then treated with cycloheximide (CHX)-mediated for 8 h. Western blot analysis of YTHDC1 protein levels over time in response to DNA damage. Data are expressed as mean ± SEM. **P < 0.01; ***P < 0.001