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. 2014 Nov 6;10(11):e1004477. doi: 10.1371/journal.ppat.1004477

Figure 10. Lipid rafts perturbation by the cholesterol-binding filipin III blocks the innate response to HSV.

Figure 10

(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 Fig. 1 C, or for phospho-Akt and total Akt (panel C), as detailed in the legend to Fig. 5. (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 Fig. 1 D. 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 Fig. 1A.