LANA promotes KSHV gene expression in 293 cells.

A-H Ratios of viral RNA to DNA following KSHV or KSHVΔLANA infection of 293 cells for the indicated viral genes at 24, 48 or 72 hpi. ns, p>0.05; *, p<0.05; **, p<0.01; ***, p<0.001;****, p<0.0001 by unpaired t test. I. Immunoblot of the indicated gene products following KSHV or KSHVΔLANA infection of 293 cells at 24 or 48 hpi at indicated MOI’s. The multiple LANA bands, which are more prominent at 48 hpi, are due to alternative initiation of translation and an alternative poly A signal.[71,72] GFP is expressed from the EF1α promoter in the recombinant viral genome. J. Gardella gel is shown at left. Phase microscopy (right, top) or detection of GFP expression by fluorescence microscopy (right, bottom) following infection of KSHV or KSHVΔLANA at 24 hpi. Cells were detected using the 4x objective on an EVOS M7000 imaging system (ThermoFisher). K. 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 293 cells. Three independent biological replicates are shown for each time point. Early lytic, late lytic, or latent classes of genes are indicated at left.