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Status of activating phosphorylations in HEK and BVrS6K1. HA-S6Kα1 immuno-precipitated from HEK 293 cells or Sf9 cells treated with or without rapamycin (50nM), were subjected to immune-complex kinase assay using GST-S6 as substrate and separated on 12% SDS-PAGE gel for autoradiography. Alternatively immune complexes were transferred on to a PVDF membrane for probing with indicated antibodies. The results represent typical of three independent observations.
