Fig. 1.

Pharmacological inhibition or depletion of mammalian target of rapamycin (mTOR) attenuates TNF-α-induced VCAM-1 expression and monocyte adhesion onto human aortic endothelial cells (HAECs). A: HAECs were treated with TNF-α at the indicated dose for 4 h followed by flow cytometric analysis of VCAM-1 surface expression (n = 3). B and C: after 16-h pretreatment with torin 1 (B) or rapamycin (rapa; C) at the indicated concentrations, HAECs were stimulated with 0.1 ng/ml TNF-α for 4 h followed by flow cytometric analysis of VCAM-1 surface expression (n = 4). D: HAECs were transfected with MTOR-targeted siRNA (siMTOR) and stimulated with TNF-α for 4 h followed by flow cytometric analysis of VCAM-1 expression (n = 6). E: HAECs were pretreated with 75 nM torin 1 or 25 nM rapamycin or transfected with MTOR-targeted siRNA and stimulated with 0.1 ng/ml TNF-α for 4 h followed by Western blot analysis of VCAM-1 expression (n = 6). Shown are representative blots. F and G: HAECs were pretreated with rapamycin or torin 1 followed by stimulation with TNF-α as described above. Adhered DiO-stained THP-1 cells were visualized by fluorescence microscopy (F) and quantified (G) (n = 4). Values are means ± SE. ctrl, control; siCtrl, control siRNA. *P ≤ 0.05 and ***P ≤ 0.0001 vs. TNF-α; ###P ≤ 0.0001 vs. ctrl (one-way repeated-measures ANOVA followed by Dunnett’s test, A–C and G; two-tailed paired t-test, D).