CRISPRi silencing of latent genes in iSLK-219 cells. (A) Region of the viral genome encoding LANA (ORF73) and position of the sgRNA. (B) Immunoblot analysis of LANA in iSLK-219 cells, and iSLK-219-dC9K cells transduced with a non-targeting (NT) sgRNA or a LANA specific sgRNA. Image representative of three biological replicates. Bars: quantification of the data. (one-way ANOVA, **** p < 0.0001). Error bars: standard deviation, ns, not significant. (C) Immunofluorescence analyses of LANA in methanol-fixed iSLK-219-dC9K cells transduced with a LANA specific sgRNA. Image representative of three biological replicates. (D) Flow cytometry analyses (MFI) of iSLK-219 (blue trace), or iSLK-219-dC9K cells transduced with a non-targeting (NT) sgRNA (yellow trace) or a LANA specific sgRNA (red trace). Note the downregulation of EGFP expression in cells where LANA has been silenced. Image representative of two biological replicates.