FIGURE 3.
Panx1 was localized to multiple sites, compartments, and microdomains. Immunolabeling of Panx1-GFP with an anti-Panx1 antibody revealed its localization at the cell surface (A, filled arrows) in a pattern that was distinct from Cx43-GFP (A, inset). Regions of interest from live BICR-M1Rk cells expressing Panx1-GFP (B, red and blue rectangles) were chosen for live imaging and imaged at t = 0, 10, 50, 70, and 110 s (C and D). Rapid time lapse imaging revealed that Panx1-GFP is distributed primarily in a uniform pattern, whereas mobile bright fluorescent clusters could be identified at the cell surface (B and C, filled arrows) and within the cell (D, unfilled arrows). Panx1-GFP was clearly localized to dynamic plasma membrane protrusions (E, arrowheads) that were evident in BICR-M1Rk cells expressing untagged Panx1 (F, arrowheads). Nuclei in A and F are stained with Hoechst 33342 (blue). Bars, 10 μm. Representative of three independent experiments.