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Fig. 2. GSK-3 is phosphorylated during mitosis and accumulates at centrosomes and spindle poles. (A) GSK-3 phosphorylation increases during mitosis. Total HeLa cell lysates were probed using antibodies against GSK-3ß and Phospho-GSK-3{alpha}/ß (Ser21/9). S, stimulated HeLa cells (see Materials and Methods); U, unsynchronised, unstimulated HeLa cells; M, mitotic HeLa cells. (B) HeLa cells stained for Phospho-GSK-3{alpha}/ß (Ser21/9) and DNA. (C) HeLa cell stained for Phospho-GSK-3{alpha}/ß (Ser21/9), total GSK-3 and DNA. (D) Localisation of phospho-GSK-3 and {gamma}-tubulin in HeLa cells treated with 1 µM nocodazole for 2 hours. Bar, 10 µm.