Protein Extraction and Western Blot Analysis

Liver samples were homogenized in lysis buffer containing 100 mM Tris, pH 8.5, 250 mM NaCl, 1 mM EDTA, 1 mM phenylmethylsulfonyl fluoride, aprotinin at 1:5000 dilution, and leupeptin at 1:5000 dilution^{46 (link)}. Tissue homogenates were centrifuged and the supernatants including tissue protein extracts were boiled in Laemmli buffer containing 10 mM dithiothreitol, then subjected to SDS-polyacrylamide gel electrophoresis. Separated proteins were transferred to nitrocellulose membranes and blocked in Tris-buffered saline containing 3% FBS. Immunoblot analyses were performed using antibodies to phospho-STAT3 (Tyr705) (#9131, Cell Signaling Technology, Danvers, MA, USA), total STAT3 (#4904, Cell Signaling Technology), phospho-STAT1 (Tyr701) (#9167, Cell Signaling Technology), phospho-STAT5 (Tyr694) (#9314, Cell Signaling Technology), FoxM1 (13147-1-AP, Proteintech, Rosemont, IL, USA), Cyclin A2 (ab181591, Abcam, Cambridge, UK), Cdk1 (#28439, Cell Signaling Technology), and PLK1 (#4535, Cell Signaling Technology) at 1:2000 dilution and actin (A2066, Sigma) at 1:5000 dilution. Quantitative data were obtained employing a ChemiDoc Touch Imaging System (Bio-Rad Laboratories, Hercules, CA, USA). Uncropped membrane images are presented in Supplementary Fig. 6.