Anti py irs1

Liver samples or hepatocyes were lysed with RIPA peptide lysis buffer (Beyotime Biotechnology, Jiangsu, China) containing 1% protease inhibitors (Pierce) and Western blotting analysis were performed according to standard procedures. Primary antibodies were used as follows: anti‐Pknox1 (1:1000, Novus, USA), anti‐IR (1:1000; Abcam, Cambridge, UK), anti‐pY‐IRS1 (1:1000; Abcam), anti‐IRS1 (1:2000; Abcam), anti‐pS‐IRS1 (1:2000; Abcam), anti‐pY‐IRS1 (1:1000; Abcam), anti‐AKT (1:1000; Cell Signaling Technology, MA, USA), anti‐pY‐AKT (1:1000; Cell Signaling Technology), and anti‐β‐actin (1:3000, Huabio, China). Protein bands were developed using the Enhanced Chemiluminescence (ECL) system and were visualized by using the ChemiScope Western Blot Imaging System (Clinx Science Instruments Co., Ltd). The gray‐scale value assay was performed by using Image J software (Rawak Software, Inc. Germany).