Fig. 2.

Internalization of the Dectin-1/glucan complex is negatively regulated by caveolin-1. Thioglycollate-elicited macrophages from Cav(−/−) and WT mice were incubated with fluorescent-labeled β-glucan (10 μg/ml) for 3 h. A and B, the cells were stained with anti-Dectin-1 antibody and analyzed for Dectin-1 (A) and glucan (B) MFI by flow cytometry. Shown are the mean ± S.E.M. of three independent experiments (n = 3). *, p < 0.05 compared with WT control. †, p < 0.05 compared with Cav(−/−) control. C, RAW cells that overexpress Dectin-1 were incubated with GP (10 mg/ml) for 0 to 180 min. The cells were lysed, and equal amounts of protein were incubated with anticaveolin-1 antibody or anti-Dectin-1 antibody. The proteins were precipitated with Protein G beads and separated by SDS-polyacrylamide gel electrophoresis. After transfer to nitrocellulose membranes, the proteins were detected by Western blot. Representative images of three independent experiments are shown. IP, immunoprecipitation.