Ab309096

The cells were lysed using Radio-Immunoprecipitation Assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with protease inhibitors cocktail (Roche Applied Science, Indianapolis, IN, USA, dilution ratio, 1:100) and phenylmethanesulfonyl fluoride (PMSF) (Roche Applied Science, Indianapolis, IN, USA, dilution ratio, 1:1000). The protein extractions were separated by 10% SDS-PAGE transferred to 0.45 μm polyvinylidene difluoride (PVDF) membranes (Sigma Aldrich, St. Louis, MO, USA) and incubated overnight with specific primary antibody. The membrane was washed with wash buffer and probed with a secondary antibody (Beyotime Biotechnology, Shanghai, China). The autoradiograms were quantified by a developing instrument (Bio-Rad, Universal Hood II, CA, USA). The information of primary antibody is as follows: NNMT antibody (1:1000, ab119758, abcam, UK); E-cadherin antibody (1:1000, ab40772, abcam, UK); N-cadherin antibody (1:1000, ab76011, abcam, UK); β-catenin antibody (1:1000, ab32572, abcam, UK); METTL14 antibody (1:1000, ab309096, abcam, UK); IGF2BP1 antibody (1:1000, #8482, Cell Signaling Technology, USA). Results were normalized to the expression of β-actin. All blots originate from the same experiment and have undergone parallel processing. All uncropped blots were included in Supplementary Fig. 8.

Western blotting analysis was performed as previously reported [19 (link)]. Antibodies used included primary antibodies against CD63 (ab134045, Abcam), TSG101 (ab125011, Abcam), WTAP (ab195380, Abcam), METTL14 (ab309096, Abcam), METTL3 (ab195352, Abcam), SOX9 (ab26414, Abcam), Ki67 (ab16667, Abcam), PCNA (#13,110, CST), BAX (#2772, CST), Cleaved Caspase-3 (#9664, CST), Cleaved Caspase-9 (#9509, CST) and GAPDH (ab8245, Abcam).