Fig. 3: PFDN mediates IFN-β- and RV-induced ISG production.

a HEK293 cells (WT, PFDN4 KO, and PFDN4 rescue) were stimulated with IFN-β (500 U/ml) for 24 h. The PFDN4 rescue cells were PFDN4 KO cells transduced with Flag-tagged PFDN4 (single clone). A heatmap summarizes the transcript levels of MYC, ISGs (IFITM1, IFITM2, IFITM3, MX1, OAS1, OAS2, and OAS3), IL11, and GAPDH from RNA-sequencing data. The colors represent the Log₁₀ (fold change) in gene counts identified in the RNA-sequencing experiments. b WT, PFDN3 KO, and PFDN4 KO cells were stimulated with IFN-β (500 U/ml), and ISGs (IFITM1 and MX1) mRNA level was measured at 24 hpi by RT-qPCR (left and middle panel). WT, PFDN3 KO, PFDN4 KO, and PFDN4 rescue cells were stimulated with IFN-β (500 U/ml), and the OAS3 mRNA level was measured at 24 hpi by RT-qPCR (right panel). Data represents the average of three experiments; error bars indicate SEM (two-way ANOVA with Dunnett’s multiple comparisons test). c WT, PFDN3 KO, PFDN4 KO HEK293 cells were transduced with vector or PFDN3-Flag or PFDN4-Flag plasmid for 24 h and stimulated with IFN-β (500 U/ml) for another 24 h. Total cell lysates were harvested and examined by western blot with indicated antibodies. d WT, PFDN3 KO, and PFDN4 KO HEK293 cells were infected with or without RV (RRV strain) at an MOI of 5 for 24 h. MX1 mRNA level was measured at 24 hpi by RT-qPCR, which was normalized to GAPDH. Data represents the average of three experiments; error bars indicate SEM (two-way ANOVA with Šídák’s multiple comparisons test). For all figures, experiments were repeated at least three times.