Gene delivery of hypoxia-inducible factor 1 alpha (HIF-1a) into skeletal muscle reduces myocardial infarct size after ischemia-reperfusion injury. We investigated early signaling of protection. DNA for HIF-1a was injected into murine quadriceps muscle followed by electroporation with the same volume of saline as sham-treatment. Muscles and hearts were sampled for Western blot in untreated animals (day 0), and 1, 3, 5 and 7 days after gene delivery or sham-treatment. Expression of the cell survival factors heme oxygenase 1 (Hmox-1), p38 and p42/44 mitogen activated protein kinases, AMP-activated protein kinase (AMPK) and Akt was tested. Hmox-1 was upregulated in muscles for 7 days after electroporation, but more in HIF-1a- than sham- treated muscles. Unlike in shams, p38 was phosphorylated in the skeletal muscle 1 day after HIF-1a gene delivery. In the heart p38 was downregulated 1 day after treatment in both groups. Sham-treatment caused phosphorylation of p42/44 in the muscle, with no increase after HIF- 1a, and no increase in the heart. AMPK was phosphorylated on day 1 in muscles and for 3 days in hearts after HIF-1a-, while unchanged after sham-treatment. Akt was phosphorylated in the muscle of both groups, with a cardiac phosphorylation in HIF-1a-treated animals which lasted until day 7. In summary, injection of saline or DNA for HIF-1a time-dependently changes the phosphorylation of various kinases, while Hmox-1 is induced by both treatments. Activation of prosurvival kinases in hearts of HIF-1a-treated animals may play a role in cardioprotection.