Y14phosphorylation does not have a definitive role in Cav-1 distribution upon AhR expression. (A) T-FGM AhR+/+ and AhR?/? fibroblasts were treated for 1 h with the phosphatase inhibitor Na3VO4 or with solvent. Total protein extracts were analyzed by immunoblotting with specific antibodies against total and phosphorylated Cav-1-Y14 protein. (B) T-FGM cells of both genotypes were treated with Na3VO4 or with solvent and cellular extracts analyzed for Cav-1 distribution by sucrose density gradients. Immunoblotting was used to detect Cav-1 and the cytosolic marker Gapdh. (C) Cav-1 was quantified in DRMs and in soluble membrane fractions and the results represented with respect to the total amount of Cav-1 in the gradient. A representative experiment is shown. (D) T-FGM AhR+/+ and AhR?/? fibroblasts were grown on glass coverslips and transfected with a wild type Cav-1-GFP or a Cav-1-Y14F-GFP non-phosphorylable mutant. Cav-1 distribution was analyzed by live cell microscopy using a CellR fluorescence equipment. The experiments were done in duplicate in two cultures of each cell genotype.