Effect of AZD2014 and rapamycin on mTORC1 and mTORC2 in renal tumors of Tsc2+/− mice.
(A) IHC analysis. Adjacent kidney sections prepared from 16-month-old Tsc2+/− mice treated with vehicle, AZD2014, or rapamycin were used for IHC analysis. Representative IHC-stained sections were presented to show phosphorylation of mTOR at S6 at S235/236 and Akt at S473 in renal tumors. Black lines are scale bars.
(B) Western blot analysis. Proteins were prepared from solid carcinomas dissected from Tsc2+/− mice treated for 2 months from the age of 14 months with vehicle, AZD2014, or rapamycin. Beta-actin was used as a loading control. Representative Western blots were presented to show phosphorylation of mTOR at S2448 and S2481, S6 at S235/236, 4E-BP1 at T37/46, Akt at T308 and S473, PKCα at T638, and MDM2 at S166.