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. 2023 Feb 6;14:434. doi: 10.1038/s41467-023-35898-2

Fig. 1. BAF is required for optimal KSHV reactivation from latency.

Fig. 1

iSLK.219 cells were transfected with non-targeting control (NTC) siRNA or BANF1 targeting siRNA 48 h prior to the addition of 25 ng/mL doxycycline. TREx-BCBL1-RTA cells were transduced with lentiviral shRNA for 72 h prior to the addition of 1000 ng/mL doxycycline. A Fluorescent microscopy imaging of iSLK.219 cells for RFP and GFP signal at 72 h post-doxycycline treatment. B The fluorescence was quantified by plate reader. C Cells were harvested for RNA at 72 h (iSLK.219) or 96 h (TREx-BCBL1-RTA) post-doxycycline treatment and subsequent RT-qPCR was performed to quantify expression of viral mRNA transcripts. D iSLK.219 cDNA was prepared from cells harvested at 0 h and 72 h post-doxycycline treatment. Global KSHV gene expression profiling was performed. Data shown are the Z-score of the fold change (2-ΔΔCt) over the geometric mean expression of three housekeeping genes averaged over two independent biological replicates. The heatmap was prepared using Partek Flow. E Cell lysates were prepared at 72 h (iSLK.219) or 96 h (TREx-BCBL1-RTA) post-doxycycline treatment and analyzed by western blotting with the indicated antibodies. F Cells were harvested and RNA isolated at 48 h post-siRNA transfection and RT-qPCR was subsequently performed to quantify BANF1 mRNA transcripts. G Cell lysates were prepared at 48 h post-siRNA transfection or 72 h post-shRNA transduction and analyzed by western blotting with the indicated antibody. P values are the result of two-tailed Student’s T tests unless otherwise specified. Error bars indicate the standard error of the mean of three independent biological replicates. Source data are provided as a source data file.