Fig. 3. CD interacts with and dissolves extra- and intracellular CCs.

(A, B) 1 mg CCs were incubated in 0.5 mM rhodamine labeled CD or PBS as control. (A) Representative images obtained by confocal laser reflection microscopy. Scale bar equals 20 μm. (B) Quantification of rhodamine fluorescence on CCs by flow cytometry. (C) ³H-CC were incubated in CD solutions of indicated concentrations overnight shaking at 37°C. Upon filtration through 0.22 μm filter plates radioactivity was determined in the filtrate (filterable/solubilized) and the retentate (crystalline). (D, E) iMacs loaded with 200 μg CC/1×10⁶ cells for 3 h prior to incubation with 1 mM rhodamine labeled CD. (D) Quantification of rhodamine fluorescence by flow cytometry. (E) Representative images obtained by confocal microscopy; rhodamine labeled CD (red), laser reflection signal (green). Scale bars equal 5 μm. (F) Intracellular CC dissolution in BMDMs treated with 10 mM CD or control for indicated times determined by polarization microscopy. Data are shown as mean +/− s.e.m of at least three independent experiments.