Hrp conjugated goat anti mouse and anti rabbit igg h l secondary antibodies

Cells were lysed on ice for 15 min in lysis buffer (Beyotime, China), then resolved by SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was blocked with 5% low-fat milk for 2 h at room temperature and then probed with antibodies: anti-PRRSV N-protein (1:1000), anti-β-actin (1:1000; Santa Cruz, CA, USA), anti-HMOX1 (1:1000; Proteintech, USA), and anti-human Nrf2 (1:1000; Proteintech) for 2 h at room temperature. Membranes were incubated with HRP-conjugated goat anti-mouse and anti-rabbit IgG (H–L) secondary antibodies (1:1000; Beyotime, China). Bound proteins were visualized with a Tanon 5200 chemiluminescence imaging system (Tanon, China).

Cells were lysed on ice for 10 min in lysis buffer (Beyotime, China), then resolved by 10% SDS-PAGE and transferred onto nitrocellulose membrane. The membrane was blocked with 10% low-fat milk for 2 h at room temperature and then incubated with antibodies: anti-PRV gB-protein (1B1, 1:5000), anti-GAPDH (1:5000), anti-P-STAT1 (1:5000), anti-STAT1 (1:1000), anti-p-JAK1 (1:1000), anti-JAK1 (1:1000) for 2 h at room temperature. Membranes were incubated with HRP-conjugated goat anti-mouse and anti-rabbit IgG (H–L) secondary antibodies (1:1000; Beyotime, China). The proteins on the membranes were observed using the Chemistar High-sig ECL Western blotting substrate (Tanon, China) and developed on a Tanon 5200 system (Tanon, China).