Western Blotting of RAGE and STAT3 Signaling

Western blotting was performed as previously described [22 (link)]. Anti-RAGE (1:1000, #6996S; Cell Signaling Technology, Inc., MA, USA), anti-STAT3 (1:1000, #12640; Cell Signaling Technology, Inc., MA, USA), anti-phosphorylated STAT3 (1:2000, #9145; Cell Signaling Technology, Inc., MA, USA), anti-cleaved caspase 3 (1:1000, #9664; Cell Signaling Technology, Inc., MA, USA), anti-LC3II/I (1:1000, #4108; Cell Signaling Technology, Inc., MA, USA), anti-Beclin1 (1:1000, #3495; Cell Signaling Technology, Inc., MA, USA) and anti-GADPH (1:1000, #2118; Cell Signaling Technology, Inc., MA, USA) were used as primary antibodies. The membranes were incubated with primary antibodies at 4℃ overnight, followed by incubation with HRP‐conjugated secondary antirabbit antibody (1:50,000; cat. no. BM2006; BOSTER Biological Technology Co., Ltd., Wuhan, Hubei, China) at 37 °C for 1 h after washing. anti-GADPH was used as an endogenous control for other proteins. Images were obtained using a chemiluminescent western blot scanner.