Fig. 6
Mettl14 and m6A modification inhibited the stability of Notch1. a The related genes of Wnt/β-catenin, Notch and Hedeghog signaling were analyzed in Mettl14 knockout cells and Mettl14 overexpressing cells, and the expression was shown as heatmap. b, c Notch1 expression levels in Mettl14 knockout cells (b) and Mettl14 overexpressing cells (c) were examined by Western blot. Gapdh served as a loading control. d Mettl14 knockout cells were treated with 2 μg/mL actinomycin D, and then Notch1 mRNA levels at the indicated time points were examined by Northern blot. Actb was a loading control. e Correlation of Notch1 and Mettl14 expression. The expression levels of Notch1 and Mettl14 were used for analysis. Pearson correlation coefficient (R) and P-value were calculated. f Notch1 knockout cells were generated through CRISPR/Cas9 approach and examined by Western blot. g, h) Sphere formation of Notch1 knockout cells. For G, typical images were shown in upper panels and calculated numbers were shown in lower panels. For H, spheres were detected for m6A levels, confirming the decreased m6A levels upon Mettl14 knockdown. i, j Sphere formation of Notch1 knockout cells. For I, typical images were shown in left panels and calculated numbers were shown in right panels. For J, invasive cells were detected for m6A levels, confirming the decreased m6A levels upon Mettl14 knockdown. k Mettl14 was silenced in Notch1 knockout T24 cells, followed by sphere formation assay. ***P < 0.001; ns, not significant, by two-tailed T test. At least three independent experiments were performed and got similar results