FIG 5.

Effect of MAVS and RIG-I depletion on KSHV reactivation. iSLK cells latently infected with KSHV.r219 were transfected with scrambled control, MAVS, or RIG-I shRNA constructs for 48 h. (A) Knockdown was confirmed by Western blotting. (B) At 48 h postreactivation viral transcript levels were monitored by qPCR. MAVS depletion resulted in a 7-fold increase and RIG-I depletion resulted in a 3.5-fold increase in viral genomes. (C) Reactivation was monitored by RFP expression using fluorescence microscopy. Both MAVS-depleted and RIG-I-depleted cells showed increased RFP compared to that in the scrambled control cells at 48 h postreactivation. (D) Mean RFP intensity was quantified using NIS Elements (Nikon imaging software) on the Nikon Eclipse Ti. Data are shown as the average mean intensity over three independent experiments. Both shMAVS and shRIG-I cells showed increased RFP intensity over that in shSCR cells following reactivation.