Panx1 regulates N2a cell proliferation. (A) Image from a confocal z-stack with orthogonal side-views of N2a cells overexpressing Panx1EGFP. Panx1 is highly localized to the plasma membrane, stained with wheat germ agglutinin (WGA), as well as to intracellular membranes. (B) N2a cells overexpressing Panx1 exhibited a reduced doubling time (14.7 hours) compared to control cells overexpressing EGFP (26.2 hours) ((*) P < 0.05 and (**) P < 0.01 for one-way ANOVA with post-hoc Tukey’s multiple comparison test, N = 3). (C) Treatment of transfected N2a cells with the Panx1 channel blocker, probenecid (1 mM), significantly reduced cell proliferation at 48 hours in both Panx1 and EGFP overexpressing cells ((**) P < 0.01 and (***) P < 0.001 for one-way ANOVA with post-hoc Tukey’s multiple comparison test). ANOVA, analysis of variance; Panx1, pannexin1.