FIGURE 2.

Bz-ATP stimulation induces ezrin translocation to the plasma membrane. Neuro2a cells were transfected with plasmids expressing GFP (A and B) or ezrin-GFP (C and D) or N-terminal domain (Nter) of ezrin-GFP (E and F). Cells were incubated at 37 °C in DMEM and treated with 1 mm Bz-ATP. GFP distribution was assessed by video microscopy. Pictures were taken before addition of Bz-ATP (A, C, and E) and 10 min after stimulation (B, D, and F). Neuro2a-hAPP cells, plated in a polylysine-coated Lab-Tek slide at 2 × 10⁴ cells/well, were incubated with DiI cell labeling solution for 15 min and stimulated (H) or not (G) with Bz-ATP (1 mm) for 10 min. Fixed and permeabilized cells were labeled with rabbit anti-phosphospecific ERM Abs and then with Alexa Fluor 488-conjugated goat anti-rabbit IgG (G and H). Arrows show green fluorescence corresponding to phosphospecific ERM (pERM). The inset shows a 2× amplification of the image indicated by the arrow in the right lower quadrant. The results are representative of four experiments performed on different days.