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Blocking peptide. Blocking peptide. rapamycin. rapamycin. ctrl. 6hr. 24hr. ctrl. 6hr. 24hr. (A). (B). mTOR. mTOR. rictor. rictor. raptor. raptor. IP: mTOR. Cho-CK. IP: mTOR. SCK. Cho-CK. Choi-CK. (C). SCK. JCK. ctrl 6hr 24hr. ctrl 6hr 24hr. ctrl 6hr 24hr.
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Blocking peptide Blocking peptide rapamycin rapamycin ctrl 6hr 24hr ctrl 6hr 24hr (A) (B) mTOR mTOR rictor rictor raptor raptor IP: mTOR Cho-CK IP: mTOR SCK
Cho-CK Choi-CK (C) SCK JCK ctrl6hr 24hr ctrl6hr 24hr ctrl6hr 24hr ctrl6hr 24hr p-S6 (S235/236) S6 p-4EBP1 (T37/46) 4EBP1 p-AKT (S473) AKT p-STAT3 (S727) STAT3 Supplementary Figure4. Inhibition of the assemblies and kinase activities of mTORC1 and mTORC2 by the rapamycin treatment. (A, B) Rapamycin-treated cells were immunoprecipitated with anti-endogenous mTOR antibody alone or in combination with a blocking peptide for the anti-mTOR antibody. The immunoprecipitated samples were analyzed by western blot. (C) Cells were treated with 100 ng/ml rapamycin or vehicle for the indicated time. Whole-cell lysates treated with rapamycin or vehicle were immunoblotted with the indicated antibodies.