Figure 9

TSN promotes the IFI16/Caspase-1/IL-1β pathway to inhibit PRRSV replication in PAMs. (A–D) PAMs grown in 12-well plates were not infected (A and B) or infected (C and D) with PRRSV NADC30-like (0.5 MOI) for 2 h at 37 ℃ and then cultured in fresh media containing various concentrations of TSN. Total protein was extracted from cell lysates at 12, 24 and 36 h, respectively. The expression levels of IFI16, GAPDH and viral N protein were analyzed by Western Blotting. (E). PAMs grown in 6-well plates were not infected or infected with PRRSV NADC30-like (0.5 MOI) for 2 h at 37 ℃ and then cultured in fresh medium containing various concentrations of TSN. At 24 hpi, the samples were subjected to Western blot analyses to measure the levels of IFI16, caspase-1, pro-caspase-1, GSDMD-N, GSDMD-F, IL-1β, viral N protein and GAPDH. (F-I) Cells grown in 24-well plates were infected with PRRSV NADC30-like (0.5 MOI) for 2 h at 37 ℃ and then cultured in fresh medium containing various concentrations of IL-1β. At 24 hpi, the samples were subjected to qRT-PCR (F), Western Blotting (H) and IFA (G and I). Scale bar: 100 μm. Statistical significances are denoted by *p < 0.05, **p < 0.01, and ***p < 0.001.