Fig. 2.
Immobilization of IR by Cav1 in living cells. (A) TIR-FM analyses. Cav1-GFP and IR-GFP were expressed in HEK293 cells, and time-lapse images at the cell surface were taken (SI Movie 1 for Cav1-GFP and SI Movie 2 for IR-GFP). Selected frames in the same area between 0 sec (green) and 7 sec (pseudored) and their merged images are shown. (B) FRAP analyses. Cav1-RFP or Cav1F92A/F94A-RFP was coexpressed with IR-GFP in HEK293 cells. The area at the cell surface of aggregated Cav-1 or mutant protein was identified by confocal images (Upper), and the areas were bleached. (C) The fluorescence recovery of IR-GFP expressed alone (plot 1) or coexpressed with Cav1-RFP (plot 2) or the Cav1-RFP mutant (plot 3), and the fluorescence recovery of Cav1-RFP (plot 4) or the Cav1-RFP mutant (plot 5) coexpressed with IR-GFP were measured.