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LMB inhibits mitogenic stimulation of p70s6k activation and 4E-BP1 phosphorylation. Quiescent HEK293 cells were stimulated with 15% serum for 1 h, with or without prior treatment with 10 ng/ml LMB for 12 h. Endogenous p70s6k was immunoprecipitated and kinase assays were carried out in the immune complexes. 4E-BP1 phosphorylation was analyzed by Western blotting of whole cell lysates. Hyperphosphorylation of 4E-BP1 resulted in slower mobility on 13% SDS/PAGE.
