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. 2010 May 17;120(6):2016–2029. doi: 10.1172/JCI39896

Figure 5. EPO upregulates angiogenic cytokine levels in cardiomyocytes and infarcted heart.

Figure 5

(A) Endothelial cell proliferation assay using BrdU incorporation. Cardiomyocytes were pretreated with EPO (10 U/ml) or saline for 48 hours. HUVECs were treated with EPO-pretreated conditioned medium (pre-EPO) or saline-pretreated medium with EPO (EPO). Specific inhibitors of VEGF (CBO-P11) and Ang-1 (anti–Ang-1 antibody) were added to the culture medium as indicated. Data from a representative experiment are shown (n = 5 per condition). *P < 0.05; #P < 0.01. (B) Tube formation assay. Quantification of tube length in HUVECs cultured with the conditioned medium was described in A. Data from a representative experiment are shown (n = 3 per condition). (C) Western blotting and quantification of VEGF levels from cultured cardiomyocytes treated with EPO. Time (at dose of 100 U/ml EPO) and dose (treated for 48 hours) dependency are shown (n = 3). *P < 0.05 versus control. (D) qRT-PCR analysis of Ang-1 mRNA from cultured cardiomyocytes treated with EPO (n = 4). Shown are levels of VEGF protein (E), Ang-1 mRNA (F), and protein (G) in the heart after MI. Representative Western blots and quantification are shown. WT mice were subjected to MI and treated with EPO or saline (control) (n = 4–5 for each condition).