Figure 5

AhR contributes in determining Cav-1 localization to DRM domains in directionally migrating fibroblasts. (A) T-FGM AhR+/+ and AhR?/? fibroblasts were grown to confluence on glass coverslips. Wounds were done to induce directional migration (indicated by an arrow). The expression patterns of Cav-1 and cholera toxin ? (DRM marker) were analyzed by immunofluorescence using a Fluoview F1000 confocal microscope. Cav-1 and cholera toxin ? were detected using secondary antibodies conjugated with Alexa 488 and Alexa 633, respectively. A merge of both expression profiles is shown on the right. (B) The same experiments were performed to determine the co-localization of Cav-1 with the soluble membrane marker TfR. Note that Cav-1 was detected using a secondary antibody labeled with Alexa 633 whereas TfR was bound to an Alexa 488-labelled secondary antibody. DAPI staining was used to label cell nuclei. Arrowheads indicate co-localization of Cav-1 with cholera toxin ? in T-FGM AhR+/+ cells or Cav-1 in T-FGM AhR?/? cells. The experiment was done in triplicate in two T-FGM cultures of each genotype.