Fig. 3: CIMSS inhibits HSV infection.

a HaCat (n = 4) or primary vaginal cells (n = 2) were treated with increasing doses of CIMSS (or DMSO control) and then infected with HSV-2(G). Plaques were counted by immunostaining after 48 h and are presented as the percent reduction in viral plaque numbers relative to the DMSO controls, which had ~200 plaques per well (mean ± SD; ***p < 0.001, ***p < 0.0001, ANOVA with multiple comparisons compared to DMSO control for HaCat cells). b HaCat cells were synchronously infected with HSV-2(G), treated with media containing 10 μM CIMSS or 0.1% DMSO at temperature shift, and at the indicated times post-temperature shift, extracellular virus was inactived with low pH citrate buffer and cells were overlaid with methylcellulose. Plaques were counted at 48 h (mean ± SEM, n = 2). Asterisks indicate significance at each time point by ANOVA (****p < 0.0001). c HaCat or primary vaginal epithelial cells were mock or synchronously infected with HSV-2(G) (MOI = 10 pfu/cell) and 0.1% DMSO, 10 μM CIMSS, 2 μg/ml rabbit anti-Akt or a control IgG were added at the time of temperature shift. Nuclear extracts were prepared after 1 h incubation at 37 °C and probed with antibodies for VP16, histoneH1 (nuclear protein) or Golgin-97 (cytoplasmic protein). The blot is representative of results obtained in two independent experiments; intensity of VP16 band (relative to histoneH1) is indicated below each lane.