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. 2023 Dec 4;14(6):e02651-23. doi: 10.1128/mbio.02651-23

Fig 6.

Fig 6

PRRSV GP5-induced ROS activates NLRP3 inflammasome. (A, B) iPAMs were infected with PRRSV (MOI = 1) simultaneously treated with NAC (10 mM) as indicated for 0–48 h. IL-1β (A) and IL-18 (B) in the medium were measured by ELISA. ***P < 0.001. (C, D) iPAMs were infected with PRRSV (MOI = 1) and simultaneously treated with NAC (10 mM) as indicated for 0–48 h. The mRNA levels of IL-1β (C) and IL-18 (D) were analyzed by qRT-PCR analysis. *P < 0.05, **P < 0.01, ***P < 0.001. (E) iPAMs were infected with PRRSV (MOI = 1) and simultaneously treated with NAC (10 mM) as indicated for 48 h. IL-1β, pro-IL-1β, pro-caspase-1, caspase-1 P20, and GAPDH were analyzed by immunoblotting analysis. (F) Caspase-1 activity was assessed in iPAMs infected with PRRSV (MOI = 1) and simultaneously treated with NAC (10 mM) or YVAD-CHO (5 µM) as indicated for 24 h. ***P < 0.001. ns, no significance. (G) The LDH activities were measured in the medium from iPAMs infected with PRRSV (MOI = 1) and simultaneously treated with NAC (10 mM) as indicated for 0–48 h. **P < 0.01, ***P < 0.001. (H, I) iPAMs were transfected with GP5-EGFP (0–4 µg) and simultaneously treated with NAC (10 mM) as indicated for 24 h. IL-1β (H) and IL-18 (I) in the medium were measured by ELISA. **P < 0.01, ***P < 0.001. (J, K) iPAMs were transfected with GP5-EGFP (0–4 µg) and simultaneously treated with NAC (10 mM) as indicated for 24 h. The mRNA levels of IL-1β (J) and IL-18 (K) were analyzed by qRT-PCR analysis. *P < 0.05, **P < 0.01, ***P < 0.001. (L) iPAMs were transfected with GP5-EGFP (0–4 µg) and simultaneously treated with NAC (10 mM) as indicated for 24 h. IL-1β, pro-IL-1β, pro-caspase-1, caspase-1 P20, and GAPDH were analyzed by immunoblotting analysis. (M) Caspase-1 activity was assessed in iPAMs cells transfected with GP5-EGFP (4 µg) and simultaneously treated with NAC (10 mM) or YVAD-CHO (5 µM) as indicated for 24 h. ***P < 0.001. ns, no significance. (N) The LDH activities were measured in the medium from iPAMs transfected with GP5-EGFP (0–4 µg) and simultaneously treated with NAC (10 mM) as indicated for 24 h. **P < 0.01, ***P < 0.001.