Lipid rafts perturbation by the cholesterol-binding filipin III blocks the innate response to HSV.

<p>(A, B) Effect of filipin III on NF-κB activation. (A) 293T- or sh-β3 293T cells were transfected with plasmids encoding TLR2, NF-κB-luc and Renilla luciferase, as indicated. Total cell lysates were subjected to SDS-PAGE and WB for IκB-α, or tubulin (panel B), as detailed in the legend to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004477#ppat-1004477-g001" target="_blank">Fig. 1 C</a>, or for phospho-Akt and total Akt (panel C), as detailed in the legend to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004477#ppat-1004477-g005" target="_blank">Fig. 5</a>. (D) Wt 293T (293T) or sh-β3 293T cells were transfected with TLR2 as indicated. IFN-β was quantified in the culture medium 48 h after infection, as detailed in the legend to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004477#ppat-1004477-g001" target="_blank">Fig. 1 D</a>. In A and D, each value represents the average of triplicate samples ± SD. In all panels filipin III treatment consisted of exposure of cells to the compound (1.5 µM) from 30 min prior to virus absorption till the end of virus absorption with gD^−/− virions, at 20 PFU equivalent/cell (panels A, B, D) or 60 PFU equivalent/cell (panel C). Cells were harvested at the indicated times, 6 h in panel A, 2 h in panel B; 10, 20, 30 min in panel C; 48 h in panel D. All other details as in the legend to <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004477#ppat-1004477-g001" target="_blank">Fig. 1A</a>.