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. 2015 Jan 30;6(6):4286–4298. doi: 10.18632/oncotarget.3006

Figure 3. Rapamycin-induced dephosphorylation of mSin1 is dependent on mTOR kinase activity.

Figure 3

Rh30 cells, infected with recombinant adenoviruses expressing GFP (Ad-GFP), FLAG-tagged rapamycin-resistant and kinase active mTOR (S2035T, Ad-mTOR-T), and rapamycin-resistant and kinase dead mTOR (S2035T/D2357E, Ad-mTOR-TE) (A) or with lentiviral shRNAs to GFP and mTOR (B) respectively, were serum-starved for 24 h. The cells were then pretreated with or without rapamycin (Rapa, 100 ng/ml) for 2 h, and further stimulated with or without IGF-1 (10 ng/ml) for 10 h, followed by Western blotting with indicated antibodies.