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Figure 1


Fig. 1. Expression of FGFRs and uptake of FGF-1 in COS-1 cells. (A) Untransfected COS-1 cells and COS-1 cells transfected with FGFR1, FGFR2, FGFR3, FGFR4, FGFR1ß, or FGFR2ß, respectively, were lysed and analyzed by SDS-PAGE and immunoblotting. The membrane was probed sequentially with antibodies specific for FGFR1, FGFR2, FGFR3 and FGFR4. (B) To cells transfected with FGFR as indicated, 35S-FGF-1 was allowed to bind at 4°C for 30 minutes and then the cells were washed and lysed immediately (upper panel) or further incubated at 37°C for 30 minutes to allow endocytosis of 35S-FGF-1 to take place and then washed and lysed (lower panel). 35S-FGF-1 was extracted from the lysates by binding to heparin-Sepharose and analyzed by SDS-PAGE and autoradiography. In the names of the receptors, FGFR is abbreviated to R. (C) COS-1 cells, untransfected or transfected with FGFR1, FGFR2, FGFR3, FGFR4, FGFR1ß or FGFR2ß as indicated, were incubated with Cy3-FGF-1 and heparin (except for the first image, which is without heparin) for 2 hours, fixed and analyzed by confocal fluorescence microscopy.





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