Reloading of cholesterol restores MIP-1β binding and calcium mobilization to BCD-treated cells.

(A) CEM-R5 cells were treated for the indicated times with BCD before washing with PBS. Treated cells were then reloaded with chol-BCD for 10 or 30 minutes, and the level of MIP-1β binding was determined as described in “Materials and methods.” Results are expressed as percentage of positive. The data are representative of 2 independent experiments. (B) BCD-treated CEM-R5 cells were reloaded with chol-BCD and analyzed for intracellular calcium mobilization as described in “Materials and methods.” Results are graphed as the average change in fluorescence ratio (F₃₄₀/F₃₈₀) of each 30-second interval. (C) Binding of MIP-1β was analyzed after BCD treatment followed by reloading with cholesterol (chol-BCD) or 4-cholesten-3-one (4C3O). MFI of each population is expressed on the graph.