Fig. 6. Cytokine profiling during ischemia reperfusion. Wild-type (WT) or Adora2b^−/−mice underwent 60 min ischemia followed by 120 min of reperfusion. The area at risk (AAR) was harvested, homogenized, and probed for various cytokines using a muliplex enzyme-linked immunosorbent assay (ELISA). Values were normalized to the total protein content of the tissue samples (A–D ). To quantify polymorphonuclear leukocyte (PMN) sequestration into the myocardium, an ELISA for myeloperoxidase (MPO) was performed (WT: 1.4 ± 0.2 ng/mg protein; Adora2b^−/−1.5 ± 0.1 ng/mg protein; not significant) (A ). To characterize the inflammatory response in WT and Adora2b^−/−mice, interferon-γ (Ιnf-γ), interleukin-1β (IL-1β), interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-5 (IL-5), keratinocyte-derived cytokine (KC), interleukin-10 (IL-10), and Interleukin-12 (IL-12) tissue concentrations were measured. Note:  no significant changes were found, whereas IL-2 and IL-10 were negative in both groups (data not shown) (B, C ). Tumor necrosis factor α (TNFα) content of the AAR in WT and Adora2b^−/−mice (WT: 7.5 ± 0.5 pg/mg protein; Adora2b^−/−: 39.3 pg/mg protein) (mean ± SD; n = 4 per group) (D ).