FIG. 6.
Effect of cholesterol removal on colocalization of CD4 or CXCR4 with F actin. CEM cells were left untreated (top panels) or treated with 10 mM MβCD at 37°C for 30 min (bottom panels). The cells were stained with anti-CD4 (A) or with anti-CXCR4 (B) followed by Alexa Fluor 594-conjugated secondary (Fab)2 antibodies (red) as described in Materials and Methods. Cells were then fixed, treated with 0.5% Triton X-100, labeled with Alexa Fluor 488-conjugated phalloidin (green) at 37°C for 30 min, and examined by confocal microscopy as described in Materials and Methods. Bar, 5 μm. The white color on the images indicates CD4-actin or CXCR4-actin colocalization.