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. 2021 Mar 23;296:100579. doi: 10.1016/j.jbc.2021.100579

Figure 1.

Figure 1

Host antiviral immune responses in BMDMs are modulated based on the immune evasion potential of IAV.A, immunoblot analysis of caspase-1 (CASP1) cleavage (pro-CASP1 (p45) and cleaved CASP1 (p20)), levels of NS1, NP, and GAPDH in WT and Nlrp3−/− bone marrow–derived macrophages (BMDMs) infected with influenza A virus (IAV) or IAV–ΔNS1 at MOI of 5. Representative blots (n = 4). B, ELISA measurement of IL-1β, IL-18, and IFN-β. ∗∗∗p = 0.001, ∗∗∗∗p < 0.0001 (unpaired two-sided t test; n > 3). Data are the mean ± SEM. C, immunoblot analysis of the levels of NLRP3, ASC, CASP1, phospho (P)-STAT1, STAT1, P-eIF2α, NP, NS1, and GAPDH proteins in BMDMs infected with IAV and IAV–ΔNS1. Representative blots (n = 2). D, confocal microscopy imaging of BMDMs infected with IAV or IAV–ΔNS1 (MOI 5) stained for G3BP1 to visualize stress granules and DAPI to visualize nuclei. The scale bars represent 10 μm (whole image) and 5 μm (magnified image). Representative images (n = 3). E, immunoblot analysis of CASP1 cleavage in WT and Ifnar1−/− BMDMs infected with IAV or IAV with IFN-β supplementation (MOI 5). IFN-β was added 3 h after infection. Representative blots (n = 2). F, immunoblot analysis of CASP1 cleavage in BMDMs infected with IAV–PR8 (MOI 20) followed by arsenite (Ars), IFN-β, or Ars + IFN-β treatment. Representative blots (n = 3). ΔNS1, NS1 deletion mutant; IFN, interferon; IL, interleukin; PR8, Puerto Rico/8/34.