IRF1 and NOS2 Mediate TNF-α and IFN-γ-Induced Inflammatory Cell Death
(A) Heatmap depicting the expression levels of type II IFN-responsive genes in WT BMDMs treated with IFN-γ alone or co-treated with TNF-α and IFN-γ for 16 h relative to their expression in untreated (Mock) BMDMs.
(B) Heatmap depicting the expression levels of type II IFN-responsive genes in patients with moderate, severe, and critical COVID-19 relative to their expression in healthy patients (Hadjadj et al., 2020).
(C) Real-time analysis of cell death in TNF-α and IFN-γ co-treated WT and Irf1–/– BMDMs. Representative images of cell death are shown at 0 h and after 48 h of TNF-α and IFN-γ treatment.
(D) Heatmap depicting the expression levels of the most downregulated genes in Irf1–/– BMDMs co-treated with TNF-α and IFN-γ for 16 h relative to their expression in WT treated BMDMs.
(E) Immunoblot analysis of iNOS in WT and Irf1–/– BMDMs co-treated with TNF-α and IFN-γ. Actin was used as the internal control.
(F) Expression analysis of NOS2 in patients with moderate, severe, and critical COVID-19 relative to the expression in healthy patients (Hadjadj et al., 2020).
(G) Real-time analysis of cell death in WT, Irf1–/–, and Nos2–/– BMDMs during co-treatment with TNF-α and IFN-γ. Representative images of cell death are shown at 0 h and after 48 h of TNF-α and IFN-γ treatment. Scale bar, 50 μm.
Data are representative of at least three independent experiments. ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001. Analysis was performed using the one-way ANOVA (F) or two-way ANOVA (C and G). Data are shown as mean ± SEM (C, F, and G). See also Figures S3, S4, and S5.