Figure 8

AhR is required to recruit Cav-1-containing vesicles to the membrane. (A) T-FGM AhR+/+ and AhR?/? fibroblasts were transfected with a Cav-1-GFP fusion protein and its recruitment to the membrane analyzed by fluorescence recovery after photobleaching (FRAP) using live cell microscopy. Images were taken before (pre-bleach) and up to 20 min after bleaching. The bleached area is indicated with a red square on the left panels. The intensities of Cav-1-GFP signals were analyzed in selected areas (green rectangles) and the profiles obtained are shown in the lower panels. Vertical arrows mark fluorescence recovery. (B) Fluorescence recovery for cells of each genotype was quantified with respect to zero time. (C) T-FGM AhR+/+ and AhR?/? cells were transfected with a Cav-1-GFP expression vector and the distance covered by the vesicles analyzed by live cell microscopy under controlled conditions. Vesicle movement was determined by taking one image on the Z axis for each millimeter during 20 min. Representative images correspond to the projection along the Z axis of pictures taken at one fixed time. (D) The accumulated and the Euclidean distances covered by Cav-1-GFP vesicles were measured and plotted for T-FGM AhR+/+ and AhR?/? cells. At least 15 vesicles from at least 3 different cells were analyzed. The assays were done in triplicate cultures of each cell line.