Fig. 8. EGFR treatment enhances viral infection and dampens the interferon response to ZIKV and HSV-1.

a Primary immortalised astrocyte cells were treated with 2 μM AG-1478 (EGFR inhibitor) for 16 h prior to infection with ZIKV and HSV-1. Cells were stained with Bodipy (493/503) to visualise LDs (green) and DAPI to visualise the cell nuclei (blue). ZIKV RNA was detected using anti-3G1.1 and 2G4 dsRNA antibodies and HSV-1 was detected using the anti-HSV-1 antibody (Abcam, ab9533), both viral proteins shown with red staining. Images are a representation of n = 3 independent experiments. b, c RT-qPCR was utilised to evaluate IFN-β, IFN-λ and viperin mRNA expression at 8, 24 and 48 hpi for both ZIKV at MOI 0.1 or HSV-1 at MOI 0.01. RT-PCRs were performed to detect viral nucleic acid levels of (d) ZIKV and (e) HSV-1. In (b–e) error bars, mean values ± SEM, P values were determined by unpaired two-tailed Student’s t test with a Holm-Sidak correction for multiple comparisons for 2 or more groups (greater than 300 cells; n = 3 biological replicates). Stimulated cells were statistically compared with their respective mock controls, ns = not significant. f IFN protein levels from these experiments were analysed via ELISA for IFN- β and IFN-λ protein at 16 h post infection. Error bars, mean values ± SEM, P values were determined by two-way ANOVA post-hoc pairwise comparisons with Bonferroni correction (n = 3 biological replicates), nd = not detected. Source data are provided as a Source Data file.