MCT-1 overexpression enhances AKT phosphorylation at residue of Ser473. Western blot analysis of extracts (150 μg) was performed on EC155-Vector and EC155—MCT-1 cell lines. Nitrocellulose membranes were immunoblotted by anti-V5 after rehybridization with anti—phospho-AKT (Ser473), AKT, and actin antibodies after stripping overnight with 200 mM glycine (pH 2.5). The phosphorylation level of AKT on Ser473 residue was increased significantly in MCT-1 overexpressing cells compared with the vector control.