Figure 4.

Rapamycin reduces VCAM-1 expression in vivo. (A) Mice were pretreated with vehicle or rapamycin and then injected with TNF. Aortas were harvested after 16 h and assayed by Western blotting. Lanes represent individual mice. (B) Expression of VCAM-1 mRNA (normalized to CD31) in aorta from mice treated as in A (n = 5 per group). (C) Mice were treated as in A. Harvested aortas were analyzed for VCAM-1 expression via immunofluorescence. (D) Expression of VCAM-1 mRNA (normalized to CD31) in aortas harvested from mice pretreated with vehicle, rapamycin, or rapamycin + MEK inhibitor (MEK-I; PD0325901) and then injected with TNF (n = 5 per group). (E) Mice were treated as in D. Harvested aortas were analyzed for VCAM-1 expression via immunofluorescence. (F) VCAM-1 expression in renal microvessels from mice pretreated with vehicle, rapamycin, rapamycin + MEK inhibitor, or MEK inhibitor and then injected with TNF. (G) IRF-1 expression in renal microvessels from mice pretreated with vehicle and rapamycin and then injected with TNF. (C and E–G) Representative images are shown. Bars: (C, E, and F) 25 µm; (G) 10 µm. (H) Leukocyte infiltration into renal glomeruli in mice treated as in F (n = 5 per group). Veh, vehicle. For B, D, and H, mean ± SEM is shown. For D and H, significance was determined by ANOVA with Tukey’s post-hoc test; all other data were analyzed with the Student’s t test. *, P < 0.05.