Anti p tbk1

Cellular proteins were extracted with RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors. Protein concentrations were measured by using a BCA protein assay kit (Thermo Fisher Scientific, USA). Protein samples were separated using 12.5% SDS-PAGE and transferred onto a polyvinylidenefluoride membrane (Millipore). Subsequently, membranes were blocked with 5% bovine serum albumin in PBS for 1 h and the appropriate antibodies at suitable concentrations were incubated overnight at 4 °C. The blots were then incubated with secondary antibodies at room temperature for 1 h. After extensive washing in TBST, protein bands were revealed with Super Singal West FemtoMaximum Sensitivity Substrate (Thermo Fisher Scientific, Rockford, USA) and visualized with Bio-Rad ChemiDocXRS system (Bio-Rad, USA). The primary antibodies were as follows: anti-GAPDH (Proteintech, 60004-1-lg), anti-p-ATM (Cell Signaling Technology, #5883), anti-p16 (Cell Signaling Technology, #80772), anti-p21 (Cell Signaling Technology, #2947), anti-cyclin E (Cell Signaling Technology, #4129), anti-cGAS (Cell Signaling Technology, #79978), anti-p-p53 (Cell Signaling Technology, #82530), anti-p-IRF3 (Cell Signaling Technology, #29047), anti-IRF3 (Cell Signaling Technology, #4302), anti-p-TBK1 (Cell Signaling Technology, #5483), and anti-TBK1 (Cell Signaling Technology, #3504).