Quantitative analysis of TNF-α and IL-1β protein expression in the primary cultured AMC after the hypoxic exposure coupled with PPADS or TNP-ATP treatment. A. Western blotting of TNF-α at 30 kDa, IL-1β at 17 kDa immunoreactive bands, respectively. B-C. Bar graphs showing significant changes in the optical density of TNF-α and IL-1β, respectively, following hypoxic exposure when compared with their corresponding controls (mean ± SD). *P < 0. 01 Control, PPADS, TNP-ATP groups vs. hypoxic group, ΔP < 0. 05 or ΔΔP < 0. 01 vs. TNP-ATP group. By western blotting analysis, IL-1β and TNF-α protein expression was significantly increased following hypoxia treatment. Blockade with PPADS, a blocker of P2X1-3, 5-7 resulted in decreased IL-1β and TNF-α protein expression after hypoxic exposure. A more drastic decrease in TNF-α and IL-1β protein expression was observed in hypoxic microglial cells blocked with TNP-ATP.