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Fig. 11. Integrin endocytosis in caveolin-1-knockdown cells. (A-F) Cells expressing caveolin-1 siRNA (shcav) or control cells (shluc) were incubated with 10 µg/ml TR-fibronectin overnight. Cells were washed, and then incubated for 8 hours in cell culture medium lacking fibronectin, but containing 50 µM chloroquine. Cells were stained with anti-β1 integrin antibody (FITC). Upper panels, shcav; lower panels, shluc. A,D, β1 integrin; B,E, TR-fibronectin; C,F, overlay images. (G-K) Cells expressing caveolin-1 siRNA (shcav, G,J) or control cells (shluc, H,K) were incubated with 50 µg/ml antibodies to 
5 integrin (G-I) or β1 integrin (J,K) at 4°C for 45 minutes. Cells were then processed for integrin endocytosis assay. The fluorescence intensity of endocytosed 5 integrin (G,H) was quantified using a MATLab-based program. (I) Fold change relative to the fluorescence intensity of endocytosed 5 integrin in shluc cells, which was set equal to 1 (mean ± range from two independent experiments). All images are optical sections collected from a confocal microscope. Scale bars: 10 µm.
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