Fig. 5: CAD-dependent pro-inflammatory responses in epithelial cells during viral infection.

A–C CAD-regulated response of HeLa cells to viral infection. A Secretion of IL-6. HeLa-supernatants cells were analyzed by ELISA after 16 h of infection with HSV-1 (MOI = 1) or after 24 h of infection with IAV (MOI = 10). HeLa control cells (non-targeting control gRNA) and HeLa CAD-deficient cells (CAD deletion by CRISPR/Cas9) were infected in parallel. B, C HeLa cells were infected with HSV-1 (B) or IAV (C) as in (A), and the expression of virus-response genes was determined by qPCR. Data are means/SEM from three independent experiments (two technical replicates each). Fold change of RNA-expression was calculated after normalizing the values of treatment group to uninfected controls. Statistics was performed using one-way ANOVA with Tukey’s test for multiple comparison correction. D–G Mouse airway epithelial cells (AEC) were isolated from wt or CAD-deficient mice. D Experimental setup. E Viral titers were determined in culture supernatants at the time points of infection indicated. F Relative mRNA expression of interferon stimulated genes and cytokines during viral infection. Fold change was calculated after normalizing the values of treatment group with uninfected controls. Statistical analysis was performed using two-way ANOVA, corrected for multiple comparisons by using the Sidak method. G Cytokine levels in basal media were measured by ELISA. Statistical analysis was done by unpaired t-test. Data are means of three transwell replicates. Error bars represent SEM. For all samples: p < 0,05: *, p < 0,01: **, p < 0,001: ***, p < 0,0001: ****.