Host innate-immune response genes and viral IE-gene expression have comparable kinetics and are activated by common factors.

<p>A) Induction of gene expression for the host <i>Tnf</i>, <i>Ifnb1</i> an <i>IL6</i> genes and the viral <i>Ie1</i> gene were measured by qPCR (n = 4) after infection with MCMV. <i>IL6</i> and <i>Ie1</i> were not expressed in uninfected sample, so Ct of 36 was set as arbitrary comparison point. B) Heat maps of Microarray data for representative innate immune genes after infection with MCMV (left panel) or stimulation with Poly IC (right panel) showing comparable kinetics of gene induction after stimulation for most of the genes. Gene expression was normalised per gene to average expression levels and colour scale indicates fold change over average expression. C) Left panel: BMDMs (n = 50) were incubated with TLR-agonists for TLR4 (LPS), TLR3 (Poly IC), TLR2 (Pam3CSK4), TLR7 (R848) and TLR9 (ODN1668) for 15 min prior to infection with MCMV-gLuc to stimulate TLR signalling (for x-axis categories see Fig 1D). IE-gene expression was measured at 2 hpi by determining gLuc activity. Asterisks depict statistical significant changes identified by Wilcoxon two-sample test (* = p-value ≤ 0.05, ** = p-values ≤ 0.01, ns = not significant). Right panel: Induction of <i>Il6</i>, <i>Tnf</i> and <i>Ifnb1</i> expression in BMDMs after stimulation (4h) with the TLR-agonists (for x-axis categories see Fig 1D) used in this study with average fold changes over mock (n = 4) measured by relative qPCR. Asterisks depict statistical significant changes identified by Wilcoxon two-sample test (* = p-value ≤ 0.05). D) Left panel: Raw264.7 (n = 18) were incubated with TLR agonists and infected with MCMV-gLuc as described for Fig 1C. IE-gene expression was measured at 2 hpi by determining gLuc activity. Asterisks depict statistical significant changes identified by Welch two-sample t-test (* = p-value ≤ 0.05, ** = p-values ≤ 0.01, ns = not significant). Right panel: Induction of host cytokines <i>Il6</i>, <i>Tnf</i> and <i>Ifnb1</i> in RAW264.7 (n = 4) cells was measured as described for Fig 1C. Asterisks depict statistical significant changes identified by Wilcoxon two-sample test (* = p-value ≤ 0.05, ns = not significant).