EGFP-CTD localization in focal adhesion plaques. (A) HeLa cells transiently expressing EGFP-CTD were mechanically sheared and labeled with TRITC-phalloidin (for F-actin) and anti-ERM antibody (detected with Cy5). Colocalization of all three proteins was observed in the focal adhesion plaques and remaining structures. (B) Line scan across PM ruffles of a PMA-stimulated HeLa cell showing EGFP-CTD (green), F-actin (red), and ERM (blue) distribution. The white line in the top image indicates the position of the line scan shown beneath. (C) Consecutive CLSM xz sections of the PM region of a transiently EGFP-CTD–expressing HeLa cell labeled with TRITC-phalloidin and anti-ERM antibody (Cy5). Colocalization of all three labels was observed in membrane extensions and cell substrate adhesion points, resulting in a white color.