Extended Data Fig. 8: Identification of IFN-induced antiviral factors that restrict vBcl-2-dependent capsid assembly and nuclear egress.
a, Representative confocal images of ORF65 staining in iSLK-BAC16.vBcl-2 KO cells transduced with ISG-specific shRNA as indicated. Scale bars, 10 μm. b, Quantification of the percentage of cells from (a) with cytoplasmic ORF65 staining (100 cells/experiment, n = 3). c, RT-qPCR validation of ISG knockdown in (a, n = 3). d,e, TRIM22 and MxB suppress infectious KSHV production. iSLK-BAC16.vBcl-2 KO cells were transduced with Ctrl or ISG-specific shRNA and induced with Dox/NaB for 72 h. Supernatants were harvested and infectious virus yield was determined by infecting SLK cells for 24 h. Representative GFP/BF images in SLK cells are shown (d). Scale bars, 430 μm. Virus yield was quantified as the percentage of GFP-positive SLK cells (e). DKD, double knockdown (n = 3). f, RT-qPCR analysis of TRIM22 and MxB mRNA in HCT116 cells infected with vBcl-2 WT or vBcl-2 KO KSHV at MOI = 10 for 60 h (n = 3). g-i, Confocal images of ORF65 staining in iSLK-BAC16.vBcl-2 WT/KO cells expressing TRIM22 and MxB post-reactivation. Percentage of cells with cytoplasmic ORF65 (h; n = 3, 300 cells/sample) and the percentage of cytoplasmic ORF65 per cell (i; n = 49 cells/sample) were quantified. Scale bars, 10 μm. Data in a,d,g are from one experiment that is representative of three independent experiments. Data in b,c,e,f,h represent mean ± s.d. analyzed by Student’s two-tailed t test or by one-way ANOVA followed by Tukey’s post hoc test. Scatter plots in i are analyzed with Kruskal-Wallis with post hoc Dunn’s test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ns, not significant. Exact P values are provided in Source Data Extended Data Fig. 8.
