FIG. 2.

Cholesterol depletion inhibits internalization of HRV2. (A) HeLa-H1 cells in 24-well plates were incubated for 30 min at 34°C with 400 nM bafilomycin (white) or 10 mM MβCD (black) or left untreated (gray). Cells were challenged with HRV2 at 30 PFU/cell for 20 min, washed three times with PBS, and incubated in fresh medium containing the respective drugs for 0, 1, and 3 h at 34°C prior to determination of virus titer, as described in Materials and Methods. Shown are the means and standard errors of two independent experiments. (B) Cells were treated for 30 min with 10 mM MβCD or left untreated as indicated. HRV2 was added, and incubation was continued for 20 min prior to fixation, permeabilization, and immunofluorescence detection of virus. Representative confocal images are shown. Virus is retained on the plasma membrane of MβCD-treated cells. Note that the cells tend to round up as a consequence of the MβCD treatment. Bar, 10 μm.