Figure 3.

Carnosol and curcumin inhibit mTOR activity and upregulate HO-1 expression in human DC via activation of AMPK. (A) Primary human DC were incubated with AICAR (1 mM), carnosol (10 μM), curcumin (10 μM), or a vehicle control for 1 h. Activation of AMPK was measured by Western blot. (B) Pooled data (n = 7) depicting densitometric analysis of phospho-AMPK expression relative to the loading control. (C) Primary human DC were incubated with compound C (5 μM) for 1 h prior to treatment with carnosol (10 μM) or curcumin (10 μM) for 1 h, followed by stimulation with LPS (100 ng/ml) for 1 h. Expression of phospho-S6 was determined by Western blot. (D) Primary human DC were incubated with AICAR (125–1,000 μM) for 24 h. Expression of HO-1 was detected by Western blot. (E) Primary human DC were incubated with compound C (5 μM) for 1 h prior to treatment with carnosol (10 μM) or curcumin (10 μM), or a vehicle control for 24 h. Expression of HO-1 was detected by Western blot. All blots depict an individual donor and are representative of 3–7 independent experiments. Blots shown are derived from the same gel(s); membranes were first probed for the protein of interest and then re-probed for β-actin as a loading control. Full-length blots are presented in Supplementary Figures 1, 2. Statistical significance was determined by one-way ANOVA with Dunnett's multiple comparisons post hoc test to compare treatment groups against the control group (^*p < 0.05).