FIG 3.

PRV infection enhances IL-1β transcription, and expression was dependent on the TLR2-TLR3-TRL4-TLR5-NF-κB axis. (A to D) Mouse peritoneal macrophages were primed with TLR2, TLR3, TLR4, and TLR5 inhibitor AN-3485 (1 μM) (A), MyD88 inhibitor T6167923 (20 μM) (B), TLR7 and TLR9 inhibitor Hydroxychl (20 μM) (C), or NF-κB inhibitor JSH-23 (20 μM) (D), respectively, for 12, 24, or 36 h. Cell viability was analyzed by detecting LDH release. Positive indicates the total LDH level in the cell and supernatant. (E to P) Mouse peritoneal macrophages were pretreated with AN-3485 (0.1, 0.5, or 1 μM) (E, I), T6167923 (5, 10, or 20 μM) (F, J), Hydroxychl (5, 10, or 20 μM) (G, K), or JSH-23 (2, 10, 20, or 40 μM) (H, L), respectively, for 1 h and then mock infected or infected with PRV at MOI of 1 for another 24 h. The mRNA and protein levels of pro-IL-1β were detected with qPCR and Western blot, respectively. The secretion levels of IL-1β in the cell supernatant were detected by ELISA in panels M to P. The results shown are representative of three independent experiments (one-way ANOVA) (panels A to H and M to P). **, 0.001 < P < 0.01; ***, P < 0.001; ns, no significance. All error bars show standard deviations (SDs).