Anti rabbit or anti mouse igg hrp conjugated antibody from sheep

Viral and host protein analysis were evaluated as previously described (55 (link), 59 (link)). Briefly, cell lysates were resolved on 7.5% Mini-Protean TGX SDS-PAGE gels and then transferred to polyvinylidene difluoride membranes using a Trans-Blot Turbo transfer system (Bio-Rad). Membranes were blocked with 5% (wt/vol) nonfat dry milk in TBST (TBS plus 0.1% [vol/vol] Tween 20) for 1 h and then probed with the indicated primary antibody in 3% (wt/vol) bovine serum albumin in TBST at 4°C overnight. After overnight incubation, the membranes were probed with the following secondary antibodies in 5% (wt/vol) nonfat dry milk in TBST for 1 h at room temperature: anti-rabbit or anti-mouse IgG-HRP-conjugated antibody from sheep (both 1:10,000; GE Healthcare). Proteins were visualized using ECL or SuperSignal West Femto chemiluminescence reagents (Pierce) and detected by autoradiography. The following primary antibodies were used: anti-pSTAT1 Y701 (1:1,000; Cell Signaling Technologies, 9171L), anti-STAT1 D1K9Y (1:1,000; Cell Signaling Technologies, 14994P), anti-IFITM1 (1:1,000; Invitrogen, PA5-20989), anti-SARS-CoV/CoV-2 Spike 1A9 (1:1,000; GeneTex, GTX632604), and anti-β-actin (1:1,000; Abcam, ab8227).