Fig. 3

METTL3 promotes CRC cell stemness in vitro. a, Representative images and quantification of the in vitro sphere-formation assay of METTL3 knockdown CRC cells and control cells (n = 6). Scale bar: 200 μm. b, In vitro limiting dilution assay of METTL3 knockdown and control SW620 cells. A well not containing spheres (diameter ≥ 50 μm) was defined as a non-response (n = 12). c, Representative images and quantification of invaded METTL3-knockdown and control SW620 and HCT116 cells. Scale bar: 100 μm. d, Cell viability of METTL3-knockdown SW620 cells and control SW620 cells after treatment with oxaliplatin for 48 h. e, Immunoblotting analysis of stem-like cell surface antigen (CD133, CD44, and EpCAM) in METTL3-knockdown and control SW620 and HCT116 cells. f, Real-time PCR analysis of SOX2 targets genes (CCND1, MYC, and POU5F1) in METTL3-knockdown and control SW620 and HCT116 cells. g, Immunoblotting analysis of SOX2 and METTL3 in METTL3-knockdown and control SW620 cells with or without SOX2 overexpression. h, Quantification of the in vitro sphere-formation assay of METTL3 knockdown and control SW620 cells with or without SOX2 overexpression. (n = 6). i, Cell viability of METTL3-knockdown and control SW620 cells with or without SOX2 overexpression after oxaliplatin treatment for 48 h. All data are presented as the mean ± SDs (n = 3). *P < 0.05, **P < 0.01 (Student’s t-test). β-Actin was used as the loading control. The relative expression level was normalized by β-Actin