Fig. 5
From: METTL3-mediated m6A mRNA modification of FBXW7 suppresses lung adenocarcinoma
m6A methylated motifs in the CDS promote FBXW7 translation. a m6A levels of fragmented RNA detected in wild-type or shMETTL3 HCC827 cells using MeRIP-qPCR. b m6A levels of predicted m6A sites analyzed by MeRIP-qPCR. Background noise was removed by normalizing samples to non-crosslinked inputs. Anti-IgG antibodies were used as a control. c Control shRNA and METTL3 knockdown HCC827 cells were transfected with or without PmirGLO-FBXW7–3’UTR-WT or PmirGLO-FBXW7–3’UTR-MUT. d Predicted m6A sites (yellow) and synonymous mutations (red) in the CDS sequence of FBXW7 mRNA. e Western blot analysis of FBXW7 expression in HCC827 cells co-expressing exogenous METTL3 and FBXW7-CDS-WT or FBXW7-CDS-MUTs. f qPCR analysis of immunoprecipitated m6A in HCC827 cells transfected with the indicated plasmids using FBXW7 PCR primers. g–i MeRIP-qPCR analysis of FBXW7 mRNA in HCC827 cells transfected with the indicated genes. Bar = mean ± SD. n.s., not significant; *p < 0.05, **p < 0.01