1. D4476 inhibits the phosphorylation of FOXO1a at Ser322 and Ser325, but not at Thr24 in vitro.
D4476 inhibits the phosphorylation of FOXO1a at Ser322 and Ser325, but not at Thr24 in vitro. Bacterially expressed GST‐FOXO1a (1 μM) was left unphosphorylated (−) or maximally phosphorylated for 30 min at 30°C with 1 U ml−1 PKB, 10 mM magnesium acetate and 0.1 mM ATP, followed by phosphorylation for 10 min with 30 mU ml−1 CK1 (+), in the presence of the indicated concentrations of D4476. Aliquots of the reaction were spotted onto nitrocellulose and immunoblotted with phospho‐specific antibodies recognizing phosphorylated Thr24 (pThr24), Ser322 (pSer322) and Ser325 (pSer325) and an antibody that recognizes the phosphorylated and unphosphorylated forms of FOXO1a equally well (FOXO1a). Similar results were obtained in several independent experiments.
Graham Rena et al. EMBO Rep. 2004;5:60-65
© as stated in the article, figure or figure legend