Anti prb s780

Cells were lysed in RIPA buffer containing protease inhibitors for 30 min on ice, followed by centrifugation at 15000 × g for 30 min. After SDS-PAGE, extracted proteins were transferred onto PVDF membranes (GE Healthcare) and probed with primary antibodies at 4°C overnight. The membranes were then incubated with secondary antibodies for 1 h, and the bands were captured through an ECL HRP substrate (Millipore). Anti-PRMT5 (Abcam, ab109451), anti-CDK4 (Santa Cruz Biotechnology, sc-260), anti-β-actin (Cell Signaling Technology, #4967), anti-β-tubulin (Cell Signaling Technology, #2146), anti-Flag M2 (Sigma-Aldrich, F3165), anti-HA (Biolegend, 901503), anti-6× His (Proteintech, 66005-1-Ig), anti-pRB S780 (Abcam, ab47763), anti-Mono-Methyl Arginine (Cell Signaling Technology, #8015) and anti-SDMA (Millipore, 07-413), anti-CCNE1 (Proteintech, 11554-1-AP), anti-CDC6 (Proteintech, 11640-1-AP) antibodies were purchased.

D, L-Sulforaphane-L-cysteine (SFN-Cys) was bought from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-β-actin, anti-α-tubulin and anti-Cyclin D1 were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-CDK4, anti-CDK6, anti-Rb and anti-p-Rb (S780) were purchased from Abcam (Cambridge, UK). Anti-ERK1/2 and anti-p-ERK1/2 (Thr202/Tyr204) and PD98059 were obtained from Cell Signaling Technology Inc (Danvers, MA, USA). Annexin V-FITC Apoptosis assay kit was acquired from Genstar (Beijing, China). Cell Cycle and Apoptosis Analysis Kit was obtained from Beyotime Biotechnology (Shanghai, China).