Figure 6

TSN treatment upregulates cellular IFI16 expression and suppresses PRRSV replication in Marc-145 cells. (A and B) Marc-145 cells grown in 6-well plates were infected or not infected with PRRSV NADC30-like (0.05 MOI) for 2 h at 37 °C and then cultured in fresh medium containing 1.6 μM TSN. At 4 h post infection, the samples were subjected to high-throughput RNA sequencing. (A) The clusterProfiler R package was used to test the statistical enrichment of differentially expressed genes in KEGG pathways. (B) Heat map of expressed NOD-like pathway genes modulated by 1.6 μM TSN or DMSO treatment in Marc-145 cells. Red and blue correspond to relative up- and down-regulation, respectively. (C) Cells grown in 12-well plates were infected or not infected with PRRSV NADC30-like (0.05 MOI) for 2 h at 37 °C and then cultured in fresh media containing various concentrations of TSN. At 24 hpi, the samples were subjected to qRT-PCR analysis. (D and E) Cells grown in 6-well plates were not infected (D) or infected (E) with PRRSV NADC30-like (0.05 MOI) for 2 h at 37 °C and then cultured in fresh media containing various concentrations of TSN. The samples were collected and total protein were extracted from cell lysates at 12, 24 and 36 hpi, respectively. Western blotting was used to analyze IFI16 and GAPDH expression. Statistical significances are denoted by *p < 0.05, **p < 0.01, and ***p < 0.001.