Fig. 2: IFNα-induced restriction is DAXX-dependent.

A, B Sympathetic neurons were infected with Stayput HSV-1 in the presence or absence of IFNα and depleted of DAXX using shRNAs at 5 days pre-infection using two independent shRNAs. Reactivation was quantified based on the numbers Us11-GFP expressing neurons following addition LY294002. Data represent the mean ± SEM. N = 9 biological replicates from 3 independent dissections. Statistical comparisons were made using an Ordinary one-way ANOVA with Tukey’s multiple comparison (A: *p = 0.0328; *p = 0.0267; ***p = 0.0001. B: *p = 0.0101; ****p < 0.0001). C, D Representative images of sympathetic neurons untreated or treated with 600 IU/ml of IFNα cultured in 5% oxygen. Neurons were infected with EdC/EdA-labeled HSV-1 and depleted of PML or DAXX at 5 days pre-infection. Cells were fixed at 1 day (C) and 10 days (D) post-infection. The viral genome was visualized using click chemistry, and immunofluorescence was carried out for DAXX. White arrows indicate viral genome. Scale bar, 10 μm. E, F NucSpotA was used to quantify the signal intensity of DAXX at the viral genome 1- or 10-days post-infection. Each data point represents one viral genome. Percentages indicate the proportion of genomes with NucSpotA intensity ratios above the denoted co-localization threshold (dashed line). N > 60 cells from 3 biological replicates. Statistical comparisons were made using E two-tailed Mann–Whitney (****p < 0.0001) and F Kruskal–Wallis test with Dunn’s multiple comparison (*p = 0.0122; ****p < 0.0001).