The effect of epigallocatechin gallate (EGCG) on glucose uptake was studied in L6 rat skeletal muscle cells. Glucose uptake assay revealed that EGCG increased glucose uptake >70% compared to control. EGCG-stimulated glucose uptake was blocked by LY294002, an inhibitor of phosphatidylinositol (PI) 3-kinase, which is a major regulatory molecule in glucose uptake pathways. However, AMP-activated protein kinase (AMPK), which is another crucial mediator in independent glucose uptake pathways, did not inhibit EGCG-stimulated glucose uptake by SB203585, a specific inhibitor of the AMPK downstream mediator, p38 mitogen-activated protein kinase (MAPK). We also found that EGCG increased the phosphorylation level of protein kinase B and PI 3-kinase activity, when assessed by PI 3-kinase assay, whereas no increase in the phosphorylation level of AMPK and p38 MAPK was observed. Taken together, these results suggest that EGCG might stimulate glucose uptake, not AMPK-mediated but PI 3-kinase-mediated, in skeletal muscle cells, thereby contributing to glucose homeostasis.