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. 2022 Jun 7;119(24):e2117636119. doi: 10.1073/pnas.2117636119

Fig. 4.

Fig. 4.

Casp8-deficient BMDM produce more IL-1β through intrinsic activation of inflammasome. (A) Quantitative RT-PCR analysis of transcriptional up-regulation of cytokines and chemokines in BMDM 5 h after LPS (10 ng/mL) stimulation. Data dots represent biological replicates (n = 3 mice per genotype). (B) Cytokine/chemokine protein levels in culture supernatants from WT and C8KOLysM BMDM treated with LPS for 24 h. n = 3 to 5 biological replicates per genotype. (C) Effect of caspase-1 inhibitor and nigericin on IL-1β production by LPS-activated WT and C8KOLysM macrophages. BMDM were treated with LPS (10 ng/mL), caspase-1 inhibitor YVAD (20 μM), and nigericin (10 μg/mL) as indicated for 6 h and IL-1β in the culture supernatants was measured by ELISA. n = 4 to 5 per genotype. (D) Heightened ROS production by C8KOLysM BMDM upon LPS activation and complete inhibition by antioxidant BHA. ROS production was evaluated with DCF-DA at 4 h after LPS treatment. Cotreatment with BHA (200 μM) abolished LPS-induced ROS production. n = 3 per genotype. (E) BHA completely prevented IL-1β production when measured at 24 h after LPS stimulation. n = 3 per genotype; *P < 0.05, **P < 0.0.1, ***P < 0.001.