RTA expression in trans leads to LANA promotion of lytic gene expression and virus production in SLK cells.

A. Heatmap of z score values showing relative viral gene expression for all KSHV genes following KSHV (WT) or KSHVΔLANA (ΔL) at 24 or 72 hpi in SLK cells (derived from same data as in Fig 5J), or iSLK cells that were induced for Rta expression. Three independent biological replicates are shown for each time point. Early lytic, late lytic, or latent classes of genes are indicated at left. Relative expression values were compared to the mean of all values for both SLK and iSLK for each gene, resulting in different z scores for SLK here compared to Fig 5J, which only included SLK. Gray color indicates absent values. B. Phase (top panels) or fluorescent microscopy (bottom panels) was performed to detect GFP expression from 293T cells 24 hours following incubation with supernatant from KSHV or KSHVΔLANA infected iSLK-RTA cells that had been induced for RTA expression. Cells were detected using the 4x objective on an EVOS M7000 imaging system (ThermoFisher). C. Viral titer of RTA induced, iSLK-RTA cell supernatant, 96 hpi following KSHV or KSHVΔLANA infection. GFP expression was detected by flow cytometry. D. Fold difference in KSHV genome copy number determined by qPCR of RTA induced, iSLK-RTA cell supernatant 96 hpi following KSHV or KSHVΔLANA infection.