Ab205722

The cells were washed with PBS and scraped using a cell scraper. The cells were transferred to 1.5 mL microtubes and centrifuged at 5000 g for 3 min. Cells were lysed in 100 μL of lysis buffer (50 mM Tris–HCl, pH 6.8, 2% SDS, 6% 2‐mercaptoethanol, 10% glycerol, 0.0125% bromothymol blue) and heated at 95°C for 5 min. Whole cell extracts were subjected to electrophoresis on 4%–20% Mini PROTEAN TGX gels (Bio‐Rad, 4561095) and then transferred to Trans‐Blot Turbo Transfer Pack membranes (0.2 μM PVDF, BIO‐RAD, 1704156). Membranes were blocked with 5% BSA in Tris‐Buffered Saline‐Tween, TBST 0.001% azide for 60 min and probed with primary anti‐STING (D2P2F) rabbit mAb (#13647, Cell Signaling Technology), anti‐cGAS (D1D3G) rabbit mAb (#15102, Cell Signaling Technology), anti‐EGF‐Receptor (D38B1) XP rabbit mAb (#4267), and anti‐β‐actin (13E5) rabbit mAb (#4970, Cell Signaling Technology). The membranes were then washed with TBST and incubated with a secondary donkey anti‐rabbit immunoglobulin antibody conjugated to horseradish peroxidase (HRP) (Abcam, ab205722) for 1 h. Proteins were detected using a chemiluminescence detection system (ECL Prime Western Blot Detection System; GE Healthcare, RPN2232) according to the manufacturer's instructions.