Mettl1

Proteins from GBM cells were obtained using RIPA lysis buffer (ThermoFisher, USA) containing a 1% protease and phosphate inhibitor cocktail. PVDF membranes (Bio-Rad, CA, USA) were incubated with specific antibodies after Western blotting was performed. The primary antibodies used in this study are as follows: METTL1 (Proteintech, USA) and β-actin (Cell Signaling Technology, USA).

Protein was extracted from BMMCs and engineered AML cell lines with NP-40 Lysis Buffer (Beyotime, catalog number: P0013F, China), supplemented with 1 mM PMSF (Beyotime, catalog number: ST506, China). Protein concentrations were assessed by the Enhanced BCA Protein Assay Kit (Beyotime, catalog number: P0010, China). For Western blot, 40 μg protein was separated on SDS-PAGE gels and transferred onto nitrocellulose membranes (Millipore, catalog number: HATF00010, USA). After blocking with 5% skimmed milk, the membranes were incubated with primary antibody against METTL1 (Proteintech, catalog number: 14994-1-AP, China), WDR4 (Santa Cruz, catalog number: sc-100894, USA), BCL2 (Proteintech, catalog number: 60178-1-1, China), Caspase3 (Proteintech, catalog number: 19677-1, China) and β-actin (Proteintech, catalog number: 66009-1-Ig, China) at 4 °C overnight. Whereafter, the membranes were incubated with Horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (Proteintech, catalog number: SA00001:15, China) and HRP-conjugated goat anti-mouse immunoglobulin G (Beyotime, catalog number: A0216, China) secondary antibody. Finally, Blots were scanned using Bio-Rad system (USA).