FIGURE 5.

TAK-779 induces rapid plasma membrane relocalization of CCR5 internalized by CCL5 but not by PSC-RANTES. CHO-CCR5 cells were treated with 100 nm PSC-RANTES or 100 nm CCL5 for 120 min at 37 °C to internalize CCR5. Cells were then washed and incubated in the presence or absence of TAK-779 (400 nm) at 37 °C. A, at the indicated times, the cells were washed at 4 °C, stained with phycoerythrin-conjugated 3A9, and analyzed by flow cytometry. Data shown correspond to mean cell surface CCR5 (n = 3; error bars, S.E.) expressed as a percentage of control levels and are representative of three independent experiments. B and C, at the indicated times, cells were washed, fixed, and labeled with anti-CCR5 (3A9; green) and anti-TGN38 (red). Maximum intensity projections from stacks of deconvoluted confocal images are shown. Scale bars, 10 μm. The images shown are representative of two independent experiments.