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. Author manuscript; available in PMC: 2011 Jul 20.

Published in final edited form as: Circulation. 2010 Jul 6;122(3):282–292. doi: 10.1161/CIRCULATIONAHA.110.953208

Cardiac MIF-AMPK axis in young and aged hearts. A, Quantitative PCR for MIF expression of non-perfused heart as described in the Supplemental Methods, n=4 per group, *P=0.01 vs. young. B, The relative levels of MIF of non-perfused hearts after normalization to β-tubulin, n=5 per group, *P=0.001 vs. young. C, MIF content in heart homogenates from young and aged hearts after control perfusion (baseline) or after ischemia and reperfusion (reperfusion) (upper panel), bars show the rates of coronary effluent MIF production from young and aged hearts during normal perfusion or washed out following 20 min of ischemia, MIF concentration was measured by ELISA and multiplied by the coronary flow rate to calculate the production rate, n=5 per group. *P<0.05 vs. baseline, respectively; †P=0.03 vs. young reperfusion. D, Levels of HIF-1α in ischemic area of young vs. aged hearts during sham control or in vivo regional ischemia (20 min), n=4 per group. *P<0.05 vs. control, respectively; ^‡P=0.01 vs. young control; †P=0.02 vs. young ischemia. E, The relative levels of HIF-1α and MIF proteins from young control or HIF-1α inhibitor (YC-1) treated hearts, n=3–4 per group, *P<0.01 vs. control, respectively. F, Quantitative PCR for MIF mRNA in isolated cardiomyocytes from young control or YC-1 treated hearts, n= 4 per group *P=0.01 vs. control. G, Phosphorylation of AMPK from young control or YC-1 treated hearts during ex vivo ischemia (20 min), n=6 per group, *P<0.05 vs. baseline, respectively; †P=0.01 vs. control ischemia. H, Hearts were subjected to in vivo regional ischemia (20 min)/reperfusion (4 hr), and dual staining to assess the extent of myocardial necrosis (upper panel). Bars represent the percent of infarct size to area-at-risk in young, aged and young YC-1 treated hearts (lower panel), n=4 per group, *P<0.05 vs. young, respectively; †P=0.03 vs. aged. I, Immunoblots of MIF content in heart homogenates from young control and YC-1 treated hearts after control perfusion (baseline) or after ischemia and reperfusion (reperfusion) (upper panel), bars show the rates of coronary effluent MIF production from young control or YC-1 treated hearts during normal perfusion or washed out following 20 minutes of ischemia, MIF concentration was measured by ELISA and multiplied by the coronary flow rate to calculate the production rate, n=4 per group, *P=0.01 vs. baseline; †P=0.03 vs. control reperfusion.

Cardiac MIF-AMPK axis in young and aged hearts. A, Quantitative PCR for MIF expression of non-perfused heart as described in the Supplemental Methods, n=4 per group, *P=0.01 vs. young. B, The relative levels of MIF of non-perfused hearts after normalization to β-tubulin, n=5 per group, *P=0.001 vs. young. C, MIF content in heart homogenates from young and aged hearts after control perfusion (baseline) or after ischemia and reperfusion (reperfusion) (upper panel), bars show the rates of coronary effluent MIF production from young and aged hearts during normal perfusion or washed out following 20 min of ischemia, MIF concentration was measured by ELISA and multiplied by the coronary flow rate to calculate the production rate, n=5 per group. *P<0.05 vs. baseline, respectively; †P=0.03 vs. young reperfusion. D, Levels of HIF-1α in ischemic area of young vs. aged hearts during sham control or in vivo regional ischemia (20 min), n=4 per group. *P<0.05 vs. control, respectively; ^‡P=0.01 vs. young control; †P=0.02 vs. young ischemia. E, The relative levels of HIF-1α and MIF proteins from young control or HIF-1α inhibitor (YC-1) treated hearts, n=3–4 per group, *P<0.01 vs. control, respectively. F, Quantitative PCR for MIF mRNA in isolated cardiomyocytes from young control or YC-1 treated hearts, n= 4 per group *P=0.01 vs. control. G, Phosphorylation of AMPK from young control or YC-1 treated hearts during ex vivo ischemia (20 min), n=6 per group, *P<0.05 vs. baseline, respectively; †P=0.01 vs. control ischemia. H, Hearts were subjected to in vivo regional ischemia (20 min)/reperfusion (4 hr), and dual staining to assess the extent of myocardial necrosis (upper panel). Bars represent the percent of infarct size to area-at-risk in young, aged and young YC-1 treated hearts (lower panel), n=4 per group, *P<0.05 vs. young, respectively; †P=0.03 vs. aged. I, Immunoblots of MIF content in heart homogenates from young control and YC-1 treated hearts after control perfusion (baseline) or after ischemia and reperfusion (reperfusion) (upper panel), bars show the rates of coronary effluent MIF production from young control or YC-1 treated hearts during normal perfusion or washed out following 20 minutes of ischemia, MIF concentration was measured by ELISA and multiplied by the coronary flow rate to calculate the production rate, n=4 per group, *P=0.01 vs. baseline; †P=0.03 vs. control reperfusion.