Fig. 4.

Knock-down of METTL3 inhibits PCa progression in vitro. a The knock-down efficiency of METTL3 shRNAs (shMETTL3–1, shMETTL3–2) with lentivirus constructs in DU-145 and PC-3 cell lines were detected by western blot assay. GAPDH was the internal reference. b and c m⁶A RNA dot-blot assay and IF were used to evaluate the m⁶A level alterations after knocking down METTL3. Methylene blue staining was loading control. d and e The proliferation ability was evaluated by colony formation assay after knocking down METTL3 (representative wells were presented) and statistically analyzed by Mann-Whitney test. f Flow cytometry assay and western blot assay were used to evaluate the apoptosis analysis induced by knock-down of METTL3. GAPDH was the internal reference. Student’s t test was used for the statistical analysis. g and h The trans-well assay and wound-healing assay (representative wells were presented) were used to determine the cell migration after knocking down METTL3. Mann-Whitney test was used for the statistical analysis. i The EMT-associated proteins and AKT phosphorylation were detected by western blot assay. GAPDH was the internal reference. Error bars represent the SD obtained from at least three independent experiments; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001