Ab51500

BAF53A: LifeSpan Biosciences LS-C196606 (Western). EZH2: Millipore 07-400 (ChIP, IP, Western). INO80: Abcam ab105451 (ChIP, IP, Western). MCRS1: Proteintech 11362-1-AP (ChIP). RUVBL1: Bethyl A304-716A (ChIP) and Abcam ab51500 (Western). RUVBL2: Bethyl A302-537A (ChIP) and Abcam ab89942 (Western). SUZ12: Cell Signaling 3737S (Western). YY1: Santa Cruz sc-1703 (Western).

The total protein was extracted from cells using RIPA lysis (Beyotime Biotechnology, Shanghai, China) containing PMSF (Thermo Fisher Scientific), and the protein concentration was determined using the BCA kit (20201ES76, Yeasen Biotechnology Co., Ltd., Shanghai, China). Quantification was performed according to different concentrations, and the protein was transferred onto a PVDF membrane (Millipore, Billerica, MA) through polyacrylamide gel electrophoresis. The membrane was blocked in 5% bovine serum albumin for 1 h at room temperature and incubated with primary antibodies against RUVBL1 (ab51500, 1: 100, Abcam, Cambridge, UK), CTNNB1 (ab273712, 1: 1000, Abcam), CDK2 (ab32147, 1: 1000, Abcam), CCND1 (ab16663, 1: 200, Abcam) at 4 °C overnight. The membrane was re-probed with horseradish peroxidase-labeled goat anti-rabbit IgG (ab6721, 1: 5000, Abcam) or goat anti-mouse IgG (ab6789, 1: 5000, Abcam) for 1 h at room temperature. The luminescent solution was added for development. Protein quantification analysis was performed by ImageJ software (National Institutes of Health) as normalized to GAPDH.