Fig. 5.

Inhibition of mammalian target of rapamycin (mTOR) is not equivalent to disruption of mTOR complex 1 and/or complex 2 in affecting human aortic endothelial cell (HAEC) signaling. A and B: HAECs were transfected with siRNA and treated with or without TNF-α for 4 h, and cell lysates were analyzed by Western blot analysis for kinase phosphorylation (n = 5). C: PKC-α phosphorylation was detected and quantified in siRNA-transfected HAECs (n = 5). D: HAECs were treated with rapamycin (rapa) and torin 1 followed by stimulation with TNF-α. Cell lysates were analyzed by Western blot analysis for PKC-α phosphorylation (n = 5). 4EBP1, 4E-binding protein-1; p-, phosphorylated; p70S6K, 70-kDa ribosomal protein S6 kinase; siCtrl, control siRNA; siMTOR, mTOR-targeted siRNA; siRAP, regulatory-associated protein of mTOR (RAPTOR)-targeted siRNA; siRIC, rapamycin-insensitive companion of mTOR (RICTOR)-targeted siRNA. *P ≤ 0.05; **P ≤ 0.0001; ***P ≤ 0.0001 vs. siCtrl (one-way ANOVA followed by a Dunnett’s test).