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. 2012 Sep 4;3:1034. doi: 10.1038/ncomms2023

Figure 8. Critical role of the intestinal MC-associated ATP-P2X7 purinoceptor pathway for induction of neutrophil infiltration.

Figure 8

P2x7+/+ and P2x7−/− BM-derived MCs were treated with 0.25 mM ATP (+) or left untreated (−). (a) Production of IL-6 (left panel; isotype mAb-treated MC, closed column; 1F11 mAb-treated MC, open column; P2x7+/+, grey column; and P2x7−/−, beige column) and TNFα (right panel) in culture supernatant (P2x7+/+, closed column; P2x7−/−, open column) was determined after 24 h stimulation. ND, not detected. Data are shown as means±s.e.m. (n=3). (b) Oncostatin M mRNA expression was measured 30 min after stimulation of P2x7+/+ (closed column) and P2x7−/− (open column) MCs with ATP. Data are shown as means±s.e.m. (n=3). (c) LT C4/D4/E4 production from ATP-stimulated (+) or -unstimulated (−) P2x7+/+ (closed column) or P2x7−/− BM-derived MCs (open column) in culture supernatants was measured by using enzyme-linked immunosorbent assay (ELISA). Data are shown as means±s.e.m. (n=3). ND, not detected. (d) P2x7+/+ and P2x7−/− BM-derived MCs were stimulated with 0.5 mM ATP. Cells were fixed and stained with an anti-5LO antibody (red) and 4′,6-diamidino-2-phenyl indole (blue). Scale bar, 10 μm. Data are representative of two experiments. (e) Representative data of a chemokine gene array are shown. Increased levels of each chemokine are shown as a heat map. (f) mRNA expression of CCL2 (left), CCL7 (middle) and CXCL2 (right) was measured by using quantitative reverse transcription–PCR. Data are shown as means±s.e.m. (n=3). (g) CCL2 production was enumerated by using ELISA 24 h after stimulation of BM-derived MCs with ATP or IgE plus antigen (IgE+Ag). Isotype mAb-treated MC, closed column; 1F11 mAb-treated MC, open column; P2x7+/+ MC, grey column; and P2x7−/− MC, beige column). Data are shown as means±s.e.m. (n=3).