The phosphorylation of hNaa10p caused by activation of GSK-3 kinase is reversed by a GSK-3 inhibitor. Cells were stimulated in vivo with Wortmannin or both LiCl + Wortmannin. Cells stimulated with neither LiCl nor Wortmannin served as negative control. 50 μg of the resuspended proteins in rehydration buffer were analyzed by 2D-SDS PAGE using 13 cm IPG pH 4–7 for IEF, and hNaa10p was detected by immunoblotting using anti-hNaa10p. The molecular size markers are indicated to the left. (A) negative control, cells were unstimulated, (B) cells were stimulated by Wortmannin in vivo (1 μM/8 hours), and (C) cells were stimulated with both Wortmannin (1 μM/8 hours) and LiCl (50 mM) in vivo.