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Fig. 2. The FERM domain of PTPL1 is necessary and sufficient for its apical localization and microvilli association in HeLa cells. (A-C) Full-length PTPL1, (D-F) PTPL1-FERM domain, and (G-I) PTPL1 {Delta}FERM with HA-epitope tags were transiently expressed in HeLa epithelial cells (see Fig. 1). Cells were treated for (left) indirect FITC localization of PTPL1 constructs with anti-HA antibody and (middle) F-actin with TRITC-coupled phalloidin. Images represent merges of horizontal confocal sections. Vertical xz sections at the level of the dotted lines are presented under each figure. Co-localizations of PTPL1 constructs and actin are represented by yellow. Full-length PTPL1 is (A) located at the apical face of cell membrane and (C) co-localized with F-actin at the level of dorsal microvilli (arrow) and lateral cell membranes. (F) The PTPL1-FERM domain displayed the same subcellular localization (arrow) as the full-length enzyme. (D) Cell extensions were also observed (arrowhead). On the contrary, (G) PTPL1 {Delta}FERM was found mainly in the cytosol and enriched around the nucleus (arrows). Bars, 20 µm.