This study determined role of PKC-α and associated inducible heat shock protein70 (iHSP70) in the repair of mitochondrial function in renal proximal tubular cells (RPTC) following oxidant injury. Wild-type PKC-α (wtPKC-α) and inactive PKC-α (dnPKC-α) mutant were overexpressed in primary cultures of RPTC and iHSP70 levels and RPTC regeneration were assessed following treatment with oxidant, tert-butylhydroperoxide (TBHP). TBHP exposure increased ROS production and induced RPTC death that was prevented by ferrostatin and necrostatin-1, but not cyclosporin A. Overexpressing wtPKC-α maintained mitochondrial levels of active PKC-α, reduced cell death, and accelerated proliferation without altering ROS production in TBHP-injured RPTC. In contrast, dnPKC-α blocked proliferation and monolayer regeneration. Co-immunoprecipitation and proteomic analysis demonstrated association between inactive, but not active, PKC-α and iHSP70 in mitochondria. Mitochondrial iHSP70 levels increased as levels of active PKC-α decreased after injury. Overexpressing dnPKC-α augmented whereas overexpressing wtPKC-α abrogated oxidant-induced increases in mitochondrial iHSP70 levels. iHSP70 overexpression: 1) maintained mitochondrial levels of phosphorylated-PKC-α, 2) improved recovery of state 3 respiration and ATP content, 3) decreased RPTC death (an effect abrogated by cyclosporin A), and 4) accelerated proliferation following oxidant injury. In contrast, iHSP70 inhibition blocked recovery of ATP content and exacerbated RPTC death. Inhibition of PKC-α in RPTC overexpressing iHSP70 blocked protective effects of iHSP70. We conclude that active PKC-α maintains mitochondrial function and decreases cell death following oxidant injury. iHSP70 is recruited to mitochondria in response to PKC-α dephosphorylation, associates with and re-activates inactive PKC-α, which promotes recovery of mitochondrial function, decreases RPTC death, and improves regeneration.