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Figure 3


Fig. 3. The interaction between GDP-Rab27a and coronin 3 is essential for endocytosis. (A) Coronin-3-silenced MIN6 cells were analyzed by immunoblotting with anti-coronin 3 and anti-Rab27a antibodies. (B) Coronin-3-silenced MIN6 cells that express GFP (green in overlay) as a transfection marker were labeled with FM4-64 (top panel, and red in overlay). Fluorescence intensity of FM4-64 on the line is shown in lower panels. Scale bar, 10 µm. (C) GFP-coronin three mutants expressing MIN6 cells were labeled with FM4-64. For rescue experiments, T7-Rab27a-T23N was co-transfected. Dashed outlines indicate transfected cells. Scale bar, 10 µm. (D) Fluorescence intensity of FM4-64 was analyzed. Among the transfected cells, the rate of cells with cytoplasmic distribution of fluorescence is given presented as a percentage. More than 40 randomly selected cells (more than ten cells per experiment) were examined. Data are expressed as the mean ± s.d. from four independent experiments. All the experiments were carried out in the presence of 25 mM glucose.





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