FIG 3.

USP9X depletion reduces viral reactivation. (A) iSLK.219 cells were either transfected with nonspecific siRNA (si NS) or USP9X siRNAs for 48 h and then treated with 1 μg/mL doxycycline for 72 h to induce lytic reactivation. Cells were harvested 72 h postreactivation and USP9X knockdown was determined by Western blotting. (B) Fluorescent microscopy images representative of cells expressing GFP and RFP are shown at 72 h postreactivation. The scale bar represents 500 μm. (C) Images from panel B were subjected to quantification of intensities of GFP and RFP channels by LAS X software. (D) Total live iSLK.219 cells treated as in panel A, were counted by trypan blue exclusion assay to determine cytotoxicity upon USP9X knockdown. These experiments were performed with at least three biological replicates.