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. 2015 Sep;94(9):1233–1242. doi: 10.1177/0022034515593465

Figure 3.

Figure 3.

P2X7 receptor is critical not only for extracellular adenosine triphosphate (eATP)–induced interleukin 1β (IL-1β) secretion but also for intracellular pro-IL-1β processing in Porphyromonas gingivalis–infected macrophages. (A) Pro-IL-1β and mature IL-1β detected by Western blot in cell lysates from wild-type (WT) or P2X7-/- bone marrow–derived macrophages (BMDMs) after 6 h of P. gingivalis infection with or without eATP as described in Materials and Methods. (B) Oxidized adenosine triphosphate (oATP; 500 μM) was added to WT BMDM culture for 30 min before and maintained during P. gingivalis infection for a total of 6 h. Cell lysates (upper panel) and cell supernatants (lower panel) with or without eATP as described in Materials and Methods. β-actin was used as a loading control. (C) IL-1β secretion was analyzed by ELISA in BMDMs pretreated with oATP (500 µM) and then infected with P. gingivalis 381 or DPG3 from WT mice. Culture supernatants were collected after 6 h of infection with P. gingivalis (multiplicity of infection, 100) and subsequent incubation with or without 5mM eATP for 30 min. Data are shown as mean ± SD and are representative of 3 independent experiments. *P < 0.05, ***P < 0.001. Protein levels were determined by densitometric quantification relative to β-actin as a loading control.