FIGURE 5.
Delivery of Panx3-GFP to the cell surface. Wild-type BICR-M1Rk cells engineered to express Panx3-GFP (A) or both Panx3 and Panx3-GFP (B) were immunolabeled with anti-Panx3 antibody. Panx3-GFP was retained mainly in an ER-like pattern (A and B, unfilled arrows) with some evidence of a cell surface distribution (A and B, filled arrows), whereas co-expression of Panx3 appeared to increase the cell surface population of Panx3-GFP (B). Rapid time lapse imaging of live BICR-M1Rk cells co-expressing Panx3-GFP and Panx3 revealed that Panx3-GFP was distributed primarily in a uniform pattern with notable mobile fluorescent clusters at the cell surface (C, filled arrows) and within the cell (C, unfilled arrow). Localization of Panx3-GFP to plasma membrane protrusions (D, arrows) was similar to that found in cells expressing only Panx3 (E, arrows). Nuclei in A, B, and E are stained with Hoechst 33342 (blue). Bars, 10 μm. Results shown are representative of three independent experiments.