Gtx122148

The cells were harvested using a curet and centrifuged at 1000× g for 10 min at 4 °C and then lysed in ice-cold radioimmunoprecipitation assay (RIPA) lysis buffer (Catalog number: 89900, Thermo Scientific company, Waltham, MA, USA) with 100 µL protease inhibitor cocktail (Roche, Pleasanton, CA, USA). Equal amounts of protein (30 µg) were separated by SDS-PAGE (10% gel) and subsequently transferred to a polyvinylidene difluoride membrane. Subsequent to blocking with 5% skimmed milk at room temperature for 1 h, the membranes were incubated at 4 °C overnight with primary antibodies, including anti-MAT2A (1:1000; GTX50027; GeneTex), anti-β-actin (1:500; tcea13161; TAICLONE BIOTECH CORP.), anti-α-tubulin (1:500; GTX112535; GeneTex), anti-Histone H3 (1:1,000; #3932; GTX122148; GeneTex), followed by incubation at room temperature for 2 h with HRP-conjugated polyclonal secondary antibody (1:5000; GTX213110-01/GTX213111-01; GeneTex). All Western blots were visualized using the enhanced plus chemiluminescence assay kit (EMD Millipore, Billerica, MA, USA), according to the manufacturer’s protocol. Protein expression levels in cells were quantified by ImageJ software (https://imagej.nih.gov/ij/, accessed on 1 May 2021).