Figure 1

Comparative activity and rapamycin response of BVr and HEK S6K. Sf9 cells infected with HA-S6K1α1 virus at an M.O.I of ≤ 1 in TNM-FH media supplemented with 5% FBS and HEK-293 cells transiently transfected with HA-S6K1 in DMEM containing 5% FBS were grown for 24 and 48 hours respectively. Cells were exposed to rapamycin or ethanol for 15 min at indicated concentrations before harvest. Lysates were clarified and used for inmunoprecipitation with α HA antibody. Immune complexes recovered were subjected to kinase assays in presence of ³²pATP using GST-S6 or S6 peptide as a substrate. Samples were separated on a 12% SDS-PAGE gel transferred on to a PVDF membrane for autoradiography or immunoblotting. Peptide kinase assays were performed by standard procedure and the activity in Cpm (Counts per minute) normalized to 100% for comparison. The data is a representative expression of four independent experiments.