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4E-BP1 and 4E-BP2 32P incorporation is increased in 293 MyrAkt cells and is resistant to wortmannin treatment. Cells were labeled with [32P]orthophosphate as described in Materials and Methods and 4E-BP1 (A) and 4E-BP2 (B) were immunoprecipitated successively with polyclonal antibodies, separated by SDS-PAGE, transferred to Immobilon-PSQ and subjected to autoradiography. Different phosphorylated isoforms are indicated for 4E-BP1.
