FIG 4.

HPgV productively infects human astrocytes and microglia. Human astrocytes (A), microglia (B), neurons (C), and a human oligodendrocyte cell line (MO3.13 cells) (D) were infected with HPgV WT or ΔNS2 (MOI, 0.1), and supernatants were harvested after 12, 24, 48, 72, and 96 h as well as 7 and 14 days postinfection. Cell supernatants were analyzed to determine the number of HPgV copies present in supernatants. (E) Cell lysates from the human astrocytes and microglia were harvested for RNA and the number of intracellular HPgV copies was measured by ddPCR and normalized to the input of 1 μg of cellular RNA. Astrocytes were infected with either HPgV WT or ΔNS2 (MOI, 0.1) for 24 h, 48 h, 72 h, 96 h, or 7 days. At each time point postinfection, cells were harvested and RNA was collected. ddPCR analysis was performed using an input of 1 μg of cellular RNA to determine the number of positive (F)- or negative (G)-strand viral RNA copies intracellularly at each time point. (*, P < 0.05; ***, P < 0.001; ****, P < 0.0001; one-way ANOVA.)