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In vitro phosphorylation of GSK-3. COS cells were treated with either insulin (1 μM) or forskolin (10 μM) for the indicated times, and PKB was immunoprecipitated. At the same time, GSK-3 was immunoprecipitated from nonstimulated cells. PKB and GSK-3 immunoprecipitates were mixed in the presence of [γ-32P]ATP, and the samples were subjected to SDS-PAGE under reducing conditions followed by autoradiography.
