Insulin signaling includes generation of low levels of H₂O₂; however, its origin and contribution to insulin-stimulated glucose transport are unknown. We tested the impact of H₂O₂ on insulin-dependent glucose transport and GLUT4 translocation in skeletal muscle cells. H₂O₂ increased the translocation of GLUT4 with an exofacial Myc-epitope tag between the first and second transmembrane domains (GLUT4myc), an effect additive to that of insulin. The anti-oxidants N-acetyl L-cysteine and Trolox, the p47^phox–NOX2 NADPH oxidase inhibitory peptide gp91-ds-tat or p47^phox knockdown each reduced insulin-dependent GLUT4myc translocation. Importantly, gp91-ds-tat suppressed insulin-dependent H₂O₂ production. A ryanodine receptor (RyR) channel agonist stimulated GLUT4myc translocation and insulin stimulated RyR1-mediated Ca²⁺ release by promoting RyR1 S-glutathionylation. This pathway acts in parallel to insulin-mediated stimulation of inositol-1,4,5-trisphosphate (IP₃)-activated Ca²⁺ channels, in response to activation of phosphatidylinositol 3-kinase and its downstream target phospholipase C, resulting in Ca²⁺ transfer to the mitochondria. An inhibitor of IP₃ receptors, Xestospongin B, reduced both insulin-dependent IP₃ production and GLUT4myc translocation. We propose that, in addition to the canonical α,β phosphatidylinositol 3-kinase to Akt pathway, insulin engages both RyR-mediated Ca²⁺ release and IP₃-receptor-mediated mitochondrial Ca²⁺ uptake, and that these signals jointly stimulate glucose uptake.