Figure 7.

METTL3 regulates membrane localization of β-catenin via Met kinase

(A) Expression levels of β-catenin and E-Cad in membrane (Mem), cytoplasmic (Cyto), and nuclear (Nu) fractions from Con and M3^Mut/− HeLa cells were detected by western blot (left) and analyzed quantitatively (right). (B) Interactions between β-catenin and E-Cad in Con and M3^Mut/− HeLa cells were detected by IP (left) and analyzed quantitatively (right). (C) Expression levels of β-catenin and phosphorylated β-catenin in Con and M3^Mut/− HeLa cells were detected by western blot (left). Percentages of phosphorylated β-catenin were analyzed quantitatively (right). (D) Expression levels of MET mRNA in HeLa, HeLa M3^Mut/−, H460 sh-NC, H460 sh-METTL3, Huh7 sh-NC, and Huh7 sh-METTL3 cells were detected by quantitative real-time PCR. (E) Expression levels of c-Met and p-AKT S473 in HeLa, HeLa M3^Mut/−, H460 sh-NC, H460 sh-METTL3, Huh7 sh-NC, and Huh7 sh-METTL3 cells were detected by western blot (left) and analyzed quantitatively (right). (F and G) Expression levels of β-catenin in Mem and Cyto fractions from HeLa cells silencing c-Met (F) and HeLa M3^Mut/− cells overexpressing c-Met (G) were detected by western blot (left) and analyzed quantitatively (right). (H) m⁶A RIP-qPCR analysis of MET mRNA in Con or M3^Mut/− HeLa cells. (I) Expression levels of MET mRNA in Con and HeLa M3^Mut/− HeLa cells treated with 5μg/mL Act-D for the indicated time were detected by quantitative real-time PCR. (J) Expression levels of MET mRNA in HeLa cells silencing IGF2BP3 were detected by quantitative real-time PCR. (K) Con or M3^Mut/− HeLa cells silencing IGF2BP3 were treated with 5μg/mL Act-D for the indicated time. Expression levels of MET mRNA were detected by quantitative real-time PCR. (L) Mechanism of m⁶A regulating cellular localization of β-catenin. Data are presented as mean ± SD from three independent experiments. Student’s t test; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.