Hrp rb

RIPA buffer (KeyGEN Biotech.) was used to lyse the cells, and protein concentration was quantified by the BCA method. An equal amount of the extracted protein was separated by 10% SDS-PAGE and transferred onto PVDF membranes. The membranes were blocked with 3% BSA for 1 h at room temperature and then incubated with the primary antibodies overnight at 4°C. After three washes with TBST, the membranes were incubated with secondary antibodies (1:3,000) for 1 h at room temperature. The membranes were washed with TBST three times before visualization by a chemiluminescence system. Antibodies used in this work: Mouse anti-CDK2 (SC-6248, Santa Cruz Biotechnology), rabbit anti-CDK4 (SC-260, Santa Cruz Biotechnology), rabbit anti-Cyclin D1 (SC-718, Santa Cruz Biotechnology), mouse anti-c-Myc (ab32, Abcam), rabbit anti-ERK (4695S, Cell Signaling Technology), rabbit anti-P-ERK (4370S, Cell Signaling Technology), rabbit anti-AKT (4685S, Cell Signaling Technology), rabbit anti-P-AKT (4060L, Cell Signaling Technology), rabbit anti-GAPDH (47724, Santa Cruz Biotechnology), rabbit anti-cleaved-Caspase-3 (9661S, Cell Signaling Technology), rabbit anti-Caspase-3 (9662S, Cell Signaling Technology), rabbit anti-cleaved-PARP1 (CY5035, Abways Technology), mouse anti-PARP1 (SC-74469X, Santa Cruz Biotechnology), HRP-Ms (7076, Cell Signaling Technology), and HRP-Rb (7074, Cell Signaling Technology).

Western blotting and immunohistochemistry were performed according to standard procedures.⁵¹ (link) The following antibodies were used: anti-HBX (ab39716, Abcam), anti-STAT1 (9172, Cell Signaling Technology), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (sc-47724, Santa Cruz Biotechnology), anti-APOBEC3A (AP1354a, Abgent), anti-FLAG (GNI4110, GNI), horseradish peroxidase (HRP)-Mouse (Ms) (7074, Cell Signaling Technology), and HRP-Rb (7076, Cell Signaling Technology).