FIGURE 9.
F-actin binds Panx1 at the carboxyl terminus. Wild type (WT) or Panx1- or Panx1-GFP-expressing BICR-M1Rk cells were lysed and subjected to immunoprecipitation (IP) for Panx1 prior to immunoblotting (IB) the immunoprecipitates and cell lysates for Panx1 or β-actin. β-Actin co-immunoprecipitated with Panx1 and Panx1-GFP (A). Monomeric actin was polymerized into F-actin, incubated with either GST fusion protein containing the carboxyl-terminal tail of Panx1 (B) or the carboxyl-terminal tail of Panx1 alone (C) and separated into supernatant or pellet fractions (denoted by S and P, respectively) prior to immunoblotting for Panx1. Panx1 was found to co-sediment with F-actin in the pellet fractions (B and C). Parallel gels were stained with Sypro gel stain, and BSA and GST were used as controls in the co-sedimentation assays. Results shown are representative of three independent experiments.