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. Author manuscript; available in PMC: 2017 Aug 10.
Published in final edited form as: Am J Transplant. 2016 May 2;16(9):2563–2573. doi: 10.1111/ajt.13808

Figure 5. The purinergic receptor P2X7 antagonist oATP inhibits macrophage polarization to M2 cells in vitro.

Figure 5

(A) Mouse bone marrow–derived macrophages were polarized toward M1 or M2 macrophages in the presence of different concentrations of oATP for 24 h. Cells were harvested; induction of Arg-1, CD206, Fizz1, iNOS, TNF-α, and IL-6 mRNA was analyzed by quantitative reverse transcriptase polymerase chain reaction; and relative levels normalized against HPRT were shown. (B) Western blot analysis of CD206 and Arg-1 protein expression by M2 cells with or without oATP. Results shown are from one of three independent experiments. (C) FACS plot showing viability of M2 cells cultured with oATP in vitro. Live bone marrow–derived macrophages were cultured under M2 conditions with or without oATP for 24 h, stained with PI and annexin V, and assessed by FACS (*p < 0.05). Arg-1, arginase 1; Ctrl, control; FACS, fluorescence-activated cell sorting; iNOS, inducible nitric oxide synthase; oATP, oxidized adenosine triphosphate; TNF-α, tumor necrosis factor α.