Anti p smc1

The protein samples were loaded and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The electrophoresed proteins were transferred to polyvinylidene fluoride (PVDF) membranes (Merck Millipore, Darmstadt, Germany), then blocked with 5% bovine serum albumin (BSA, Gbcbio, China). Membranes were probed with anti-p-H2AX (CST, United States), anti-H2AX (CST, United States), anti-p-SMC1 (Abcam, United Kingdom), anti-SMC1 (CST, USA), or anti-βactin antibodies (Proteintech, China), followed by a goat anti-rabbit IgG secondary antibody (#SA00001-2, Proteintech, China) or goat anti-mouse IgG secondary antibody (#SA00001-1, Proteintech, China) for 1 h. The blots were visualized using an ECL chemiluminescence substrate kit (#WBKLS0100, Millipore, United States).

Cycloheximide was purchased from Cell Signaling Technology (Danvers, MA, USA). MG132 was purchased from Beyotime Biotechnology (Shanghai, China). Puromycin, doxycycline, G418 and EZview Red Anti-Flag M2 Affinity Gel was obtained from Sigma (Sigma-Aldrich). Protein A&G Agarose were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Glutathione-sepharose 4B beads were from GE Healthcare (München, Germany). Anti-K48 antibody was purchased from Cell Signaling Technology detecting the endogenous levels of ubiquitination. Anti-Flag, anti-BAF155, anti-Myc, anti-GAPDH were purchased from Cell Signaling Technology. Anti-PSMA7, anti-PSMC3, anti-PSMC1, anti-PSMA1, anti-HBx were purchased from Abcam (Cambridge, UK). HRP-coupled secondary antibodies were obtained from Santa Cruz Biotechnology.