Fig. 6: METTL3 O-GlcNAcylation enhances MCM10 mRNA stability in an m6A-IGF2BP3-dependent manner.

A, B RT-qPCR to observe the effect of METTL3 O-GlcNAcylation on relative MCM10 mRNA expression in hepatoma cells. C, D Half-life of MCM10 mRNA in Huh-7 and PLC/PRF/5 cells infected with shMETTL3 lentivirus followed by transfection with Ad-METTL3 (WT or 3A). Transcription was inhibited by actinomycin D (5 μg/mL). E, F Huh-7 and PLC/PRF/5 cells were transfected with shControl or shIGF2BP3 lentivirus. Thirty-six hours after infection, cell lysates were subjected to RT-qPCR to observe MCM10 mRNA expression. G RIP-qPCR assay of MCM10 using IgG or IGF2BP3 antibodies in Huh-7 and PLC/PRF/5 cells. H, I RIP-qPCR assay of MCM10 in Huh-7-shMETTL3-WT (or 3A mutant) and PLC/PRF/5-shMETTL3-WT (or 3A mutant) cells. Half-life of MCM10 mRNA in Huh-7 (J) and PLC/PRF/5 (K) cells infected with shIGF2BP3 lentivirus. L Western blot analysis of MCM10 levels in Huh-7 and PLC/PRF/5 cells infected with IGF2BP3 shRNA. All data represent mean ± SD from three independent experiments. Data in (A–F and H–K) were analyzed using one-way ANOVA followed by Tukey tests. Data in (G) were analyzed using an unpaired, two-tailed Student’s t-test. **P < 0.01, ***P < 0.001.