FIG. 8.

Detergent-resistant membrane fraction localization and methyl-β-cyclodextrin treatment. (A) U937 cells were incubated in the absence (a, b, c, and d) or in the presence (e, f, and g) of E recombinant protein for 1 h at 37°C and lysed with 1% Triton X-100 and subjected to flotation on a discontinuous sucrose gradient. Gradient was fractionated from the top to the bottom as indicated. Equal volumes from each fraction were analyzed by SDS-10% PAGE followed by immunoblotting with anti-HSP90 (a and e), anti-HSP70 (b and f), anti-CD14 (c), anticaveolin (d), and anti-DEN virus E protein (g). (B) Infection inhibition induced by methyl-β-cyclodextrin treatment. Human peripheral blood monocytes were treated with increased concentrations of methyl-β-cyclodextrin for 1 h at 37°C prior to DEN-2 infection at an MOI of 5. Culture supernatants were harvested after 48 h of infection and assayed to determine the infectious virus titer. Each experimental point is presented as the mean of results obtained from three separate experiments.