Fig 1: ATP release through pannexin channels required for mechanosensitive priming of IL-1ß in astrocytes. (A) Swelling rat astrocytes in hypotonic solution led to a release of ATP into the extracellular medium, as detected by the luciferin/luciferase assay. The ATP hydrolase apyrase (1 U/ml) substantially reduced the response. Symbols represent mean ± SEM, n = 10. (B) Quantification of extracellular ATP levels 18 min after exposure to solutions (*Swell vs. Control isotonic, p < 0.05, ** Swell vs Swell+Apyrase, p < 0.05, n = 10). (C) Swelling astrocytes in the presence of apyrase also prevented the rise in IL-1ß mRNA (*p = 0.02 Swell vs control, **p = 0.03 Swell vs Swell+Apyrase, n = 3). (D) The swelling-induced release of ATP was inhibited by pannexin channel blocker carbenoxolone (CBX, 10 µM, *p < 0.05, Swell vs. control, **p < 0.05 Swell vs. Swell+CBX, n = 20, normalized to swell). (E) The swelling-induced rise in IL-1ß mRNA was also inhibited by 10 µM carbenoxolone (*p < 0.05, Swell vs. control, **p < 0.05 Swell vs Swell+CBX, n = 7, normalized to swell). (F) Pannexin blocker probenecid (Prob, 1 mM) reduced the swelling-induced rise of IL-1ß in astrocytes (p = 0.029). The specific peptide blocker 10Panx1 (100 µM) reduced the expression of IL-1ß as compared to the scrambled peptide control (10panxscr, *p = 0.003). Swelling alone raised IL-1ß (p = 0.03, n = 3 for all). (G) Left: Astrocytes stained for panx1 (green), actin (red) and DAPI (blue). Right: No signal was detected in the absence of panx1 antibody.
Supplier Page from Abcam for Anti-Pannexin 1 antibody