Fig. 4

METTL3 triggered enhanced FBXW7 mRNA translation. a FBXW7 mRNA was analyzed by RT-PCR in HCC827 cells with METTL3 overexpression or depletion. b Nuclear and cytoplasmic FBXW7 mRNAs were extracted separately. c FBXW7 mRNA was analyzed in HCC827 cells transfected with METTL3 control or shRNA plasmids after ActD treatment. d Western blot analysis of the cytoplasmic and nuclear fractions of HCC827 cells using polyclonal antibodies against METTL3, METTL14, WTAP, YTHDF1/2/3, LaminB (a nuclear protein), and α-tubulin (a cytoplasmic protein). e FBXW7 protein expression was determined by western blot analysis in HCC827 cells treated with CHX to block protein synthesis. f HCC827 cells were pretreated with CHX or MG-132 and then treated with control vectors or METTL3 plasmids. g Firefly (F-Luc) values were normalized against Renilla luciferase levels, and FBXW7 translation efficiency was calculated for the pmirGLO-FBXW7 reporter relative to pmirGLO in METTL3 knockdown and control HCC827 cells. Bar = mean ± SD. n.s., not significant; **p < 0.01