Anti stat1

WCL and nuclear fraction were denatured at 95°C for 10 min and separated by 12.0% SDS-PAGE gel. Proteins were transferred onto nitrocellulose membranes and probed with anti-IRF3 (Cat.A303-384A, Bethyl Laboratories Inc), anti-IRF7 (Cat. 3941; Prosci Inc.), anti-TBK1 (Cat.3504, Cell Signaling Technology), anti-phospho-TBK1 (Ser172) (Cat.5483, Cell Signaling Technology), anti-NF-κB (Cat. 8242T, Cell Signaling Technology), anti-Ets-2 (cat. sc-365666, SCBT), anti-ETV5 (Cat. sc-100941, SCBT), anti-Stat1 (Cat. sc-591, SCBT), anti-AP-2(Cat. sc-12726, SCBT), anti-SP1(Cat. sc-17824, SCBT), anti-iNOS (Cat. 610599, BD Bioscience), anti-β-actin (Cat.4970, Cell Signaling Technology), and anti-Histone H3 (Cat.9717, Cell Signaling Technology) antibodies, followed by goat anti-rabbit IgG-HRP (Cat.31460, Thermo Scientific) or goat anti-mouse IgG-HRP (Cat.31438, Thermo Scientific).

Protein samples were lysed in 5 × SDS sample buffer containing beta-mercaptoethanol, boiled at 95°C for 10 min and loaded into each well for SDS-PAGE. Samples were then transferred to PVDF membrane (Thermo) and immunoblotted using anti-NDUFA9 (Invitrogen), anti-NDUFS3 (Invitrogen), anti-NDUFA13 (Invitrogen), anti-ATP5A (Invitrogen), anti-SDHA (CST), anti-VDAC (CST), anti-HSP60 (CST), anti-PHB1 (CST), anti-PDH (CST), anti-GAPDH (Sigma), anti-GRP78 (SCBT), anti-ABCA1 (SCBT), anti-FLOT1 (CST), anti-STAT3 (CST), anti-STAT1 (SCBT), anti-AMPK (CST), anti-ERK1/2 (CST), anti-p38 (CST), anti-SYP (Abcam), anti-ACSL4 (Abcam), all diluted in 5% BSA:TBST at 1:1000, followed by appropriate HRP-conjugated secondary antibodies (Thermo) incubation (diluted in 5% non-fat milk:TBST at 1:10,000), and developed using the SuperSignal^TM West Femto Maximum Sensitivity Substrate (Thermo).