FIG 3.

Human ANP32 proteins play a direct role in primary cRNA synthesis. ActD-treated cRNP stabilization assay in eHAP WT and dKO cells, following infection with PR8 (MOI = 3), 3 hpi. (A) Accumulation of segment 6 vRNA and +RNA in eHAP WT and dKO cells, analyzed using RNAscope. Magenta arrowheads highlight a subset of NA vRNA-stained puncta. Images are representative maximum-intensity projections. (B to D) Accumulation of segment 6 vRNA (B), cRNA (C), or mRNA (D) analyzed by RT-qPCR. The dotted line indicates the background RNA present in control samples transfected with a plasmid mix lacking PB1. The fold change was calculated versus mock-infected cells. n = 3 biological replicates, plotted as means ± the SD. Significance was assessed using an unpaired t test following log transformation. ns, not significant; ****, P < 0.0001.