Elucidation of protective mechanisms against ischemia-reperfusion injury is vital to the advancement of therapeutics for ischemic heart disease. We have previously shown that cardiac-specific overexpression of fibroblast growth factor 2 (FGF2) results in increased recovery of contractile function and decreased infarct size following ischemia-reperfusion injury and have established a role for the MAPK signaling cascade in the cardioprotective effect of FGF2. We now show an additional role for the protein kinase C (PKC) signaling cascade in the mediation of FGF2-induced cardioprotection. Overexpression of FGF2 in the heart resulted in decreased translocation of PKC delta but had no effect on PKC , , or . In addition, multiple alterations in PKC isoform translocation occur during ischemia-reperfusion injury in FGF2 transgenic hearts as assessed by Western blot analysis and confocal immunofluorescent microscopy. Treatment of FGF2 transgenic and non-transgenic hearts with the PKC inhibitor, bisindolylmaleimide (1µmol/L), revealed the necessity of PKC signaling for FGF2-induced reduction of contractile dysfunction and myocardial infarct size following ischemia-reperfusion injury. Western blot analysis of FGF2 transgenic and non-transgenic hearts subjected to ischemia-reperfusion injury in the presence of a PKC pathway inhibitor (bisindolylmaleimide, 1 µmol/L), an MEK/ERK pathway inhibitor (UO126, 2.5 µmol/L), or a p38 pathway inhibitor (SB203580, 2 µmol/L) revealed a complicated signaling network between the PKC and MAPK signaling cascades which may participate in FGF2-induced cardioprotection. Together, these data suggest that FGF2-induced cardioprotection is mediated via a PKC-dependent pathway and that the PKC and MAPK signaling cascades are integrally connected downstream of FGF2.