Figure 2.

Subcellular localization of PTEN and MAGI-2. MDCK cells were plated onto fibronectin-treated cover slips, then transfected with FLAG-PTEN or HA-MAGI-2. After 24 h the cells were washed, fixed in paraformaldehyde, permeabilized in 0.2% Triton X-100, and blocked in 3% BSA in PBS containing 1 mM CaCl₂ and 1 mM MgCl₂. Cells were incubated in primary antibody in blocking buffer for 1 h at 37°C. Cells were washed in blocking solution plus 0.2% Triton. Cells were incubated with secondary antibody in blocking buffer for 45 min, washed, and then visualized by using a Zeiss LSM310 laser scanning confocal microscope.