Figure 2. Deficiency of Caspase-6 Does Not Influence NLRP3 Priming and Viral Replication in Bone-Marrow-Derived Macrophages.

(A) Immunoblot analysis of phosphorylated IκBα (pIκBα), total IκBα (tIκBα), phosphorylated ERK (pERK), total ERK (tERK), NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), pro–IL-1β, and pro- caspase-6 (pro-CASP6) in bone-marrow-derived macrophages (BMDMs) after influenza A virus (IAV) infection at the indicated time points. Actin is used as the internal control.

(B) Immunoblot analysis of pro- and cleaved caspase-1 (CASP1) in BMDMs primed with or without Pam3CSK4 (PAM3) for 5 h and then infected with IAV for 16 h.

(C) Immunoblot analysis of phosphorylated STAT1 (pSTAT1), total STAT1 (tSTAT1), Z-DNA binding protein 1 (ZBP1), influenza non-structural protein 1 (NS1), and influenza matrix protein 1 (M1) in BMDMs after IAV infection at the indicated time points. Actin is used as the internal control.

(D) Real-time PCR analysis of IAV M1 mRNA or vRNA in BMDMs after infection at the indicated time points, presented relative to levels of the host gene actin.

(E) Endpoint replication of IAV (MOI, 10) in BMDMs for 8 h. NS, not significant; *p < 0.05; ***p < 0.001 (two-way ANOVA).

Data are shown as mean ± SEM (D and E). Data are representative of three independent experiments.