Reduction of preformed dsRNA by virion-derived vhs is sensitive to hippuristanol.

Abstract

A) HeLa cells were transfected with fwd or fwd/rev for 20 hours. Cells were then mock treated or treated with RNaseIII or RNaseA/T1 (high salt buffer) and analyzed by immunofluorescence with the dsRNA-specific antibody J2 (green). B) HeLa cells were transfected with fwd/rev for 20 hours and mock infected or infected with HSV-1 (KOS) and HSV-2 WT and Vhs- at an MOI of 20 for 5 hours in the continuous presence of actinomycin D. After immunofluorescence staining with the J2 antibody the ratio of J2 positive cells was determined by analyzing more than 100 mCherry-positive cells per coverslip. The ratio of J2 positive to mCherry positive cells in the mock infected sample was set as 100%. The graph represents the average of 4 independent experiments. P-value vs. mock: * pC) Hippuristanol (hip) antagonizes the dsRNA-destabilizing effect of vhs. The experiment was performed as described in B) with addition of vehicle control (0nM) or hip at 80 or 160nM during the course of infection. The graph represents the average of 4 independent experiments. P-values WT 0nM hip vs. mock 0nM hip: * p<0.005, ** p<0.0005; p-value 160nM hip vs. 0nM hip: p<0.05 (HSV-2), p<0.005 (HSV-1).</p