Figure 6
Phosphorylation of vinculin in TAC-treated Caco2 cells in the presence of genistein. (A) Caco2 cells in the presence of genistein were stimulated with 10−7 mol/L testosterone-HSA for the indicated time periods. Following cell lysis equal amounts of proteins were immunoprecipitated (IP) with an antiphosphotyrosine (p-Tyr) antibody. The tyrosinephosphorylated as well as equal amounts of total lysates were immunoblotted (IB) with a specific antibody against vinculin. The intensity of phosphorylated vinculin bands was normalized to the intensity of the corresponding total vinculin bands. (B) Confocal laser-scanning microscopic analysis of vinculin and actin in mAR-activated Caco2 cells in the presence of genistein. Cells treated with 10−7 mol/L testosterone-HSA for different time periods were cultured in coverslips, fixed and stained with mouse antivinculin, anti-mouse-FITC as secondary antibody, DRAQ5™ for nuclei staining and rhodamine-phalloidin for filamentous actin staining. Magnification 100×.