Western Blot Analysis of Protein Targets

Western blot analysis was conducted as previously described.^{79 (link)} In brief, total protein levels were quantified and loaded onto an SDS-PAGE gel. The separated proteins were then transferred onto a PVDF membrane (Millipore, MA, USA) via the Bio-Rad Trans-Blot Turbo™ transfer apparatus (USA). Following blocking with 5% non-fat dry milk in TBST, the membrane was incubated overnight at 4 °C with primary antibodies against FOXK1 (1:1 000, Abclonal, CHN), STAT1 (1:1 000, proteintech, CHN), STAT2 (1:1 000, proteintech, CHN), HDAC3 (1:1 000, proteintech, CHN) and β-actin (1:2 000, Abclonal, CHN). Following this, the membrane was rinsed thrice using TBS-T and then incubated with suitable HRP-linked secondary antibodies for one h at room temperature. Chemiluminescent detection was subsequently carried out using the enhanced chemiluminescence kit (ABclonal, CHN) and the blots were visualized using the Tanon Multi5200 chemiluminescence imaging system (Tanon, Multi5200, CHN).