Figure 7.
TBK1 deletion impairs BMM and AM transcriptional responses differently during influenza virus infection in vitro and has subtype-specific effects on lung macrophages in vivo. RT-qPCR of (A) AMs or (B) BMMs infected with multiplicity of infection 1 of IAV strain PR8 in vitro for 6 hours. Data were normalized to the 18S mRNA and expressed as fold change compared with uninfected WT by the comparative cycle threshold method. Heatmap depicts z-score of mRNA transcript expression level scaled to row mean. n = 3 per group and two independent experiments. Asterisks depict significance of P values for infected WT versus TBK1-KO samples (*P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001). More detailed graphs for each gene are shown in Figure E6 in the data supplement. (C) Macrophage subpopulations were isolated from lung digests using FACS at Day 4 after infection and subjected to RT-qPCR of indicated genes. mRNA levels were normalized to ActB or to 18S by the comparative cycle threshold method. Data reflect three pooled mice analyzed in technical triplicate. All rows attain statistical significance by two-way ANOVA. Symbols depict significance (P < 0.05) by Šidák’s multiple comparisons method between WT and KO for AMs#, IMs‡, or InfMsφ. Detailed graphs for each gene are shown in Figure E7.