Fig. 5.

Aloe-emodin inhibits mTORC2 kinase activity in vitro. (A) Aloe-emodin has no effect on mTOR protein abundance. The effect of aloe-emodin on mTOR kinase activity was measured by an in vitro kinase assay using [γ-³²P] ATP and a glutathione-S-transferase-Akt1 protein as substrate. ³²P-labeled glutathione-S-transferase-Akt1 was visualized by autoradiography as described in Materials and methods. (B) Aloe-emodin inhibits mTORC2 kinase activity. The effect of aloe-emodin on mTORC2 kinase activity was measured by an in vitro kinase assay using a glutathione-S-transferase-Akt1 protein. The phosphorylation level of Akt (Ser473) was visualized by western blotting.