Anti myc

All the antibodies used in the study were purchased commercially, including Anti-Flag tag (Sigma, catalog F7425), anti-Myc (HuaBio, catalog EM31105), anti-BAP1 (Santa Cruz Biotechnology, catalog sc-28383 for IP and WB; GST, catalog 78105 for ChIP), anti-ASXL1 (Abcam, catalog ab228009; Abcam, catalog ab221455), anti-FOXK1 (Abcam, catalog ab18196), anti-FOXK2 (Abcam, ab5298), anti-HCF-1 (GST, catalog 50708), anti-H2AK119 mono-ubiquitination (Abcam, catalog ab193203), anti-H3K27me3 (Abcam, catalog ab6002), anti-Histone H2A (Abcam, catalog ab18255), anti-Histone H3 (CST, catalog 9715), anti-TXNIP (Abcam, catalog ab188865), anti-HIF-1α (Abcam, catalog ab216842), anti-STAT3 (CST, catalog 12640), anti-p-STAT3 (CST, catalog 9145), anti-JAK2 (Abcam, catalog ab108596), anti-p-JAK2 (Abcam, catalog ab32101), anti-GAL4 (Santa Cruz Biotechnology, catalog sc-510), anti-ACTIN (Genscript, catalog A00702).

Total proteins were extracted using lysis buffer consisting of 2% sodium dodecyl sulfate (SDS), 1% Triton X-100, 50 mM Tris-HCl, and 150 mM NaCl (pH 7.5), separated by 10% SDS-PAGE, and transferred to a nitrocellulose membrane. The membrane was then blocked with 5% skim milk and incubated with primary antibodies at 4°C overnight. After hybridization with horseradish peroxidase (HRP)-conjugated secondary antibodies, the membrane was visualized using enhanced chemiluminescence (ECL) reagents (Bio-Rad, Hercules, CA, USA), and a Quantity One system (Bio-Rad, USA) was applied for analysis. The primary antibodies were anti-myc (Huabio, China), anti-TNFAIP3 (Proteintech, USA), anti-β-actin (Huabio, China), anti-caspase-3 (Cell Signaling Technology [CST], USA), anti-AGO2 (Abcam, USA), and anti-PDCoV N protein (Medgene Labs, USA), and the anti-PDCoV S protein polyclonal antibody was stored in our lab.