Inhibition of TRAF6 abolishes LPS-induced Akt activation. Endothelial cells transduced with a dominant-negative TRAF6 (TRAF6-C) were exposed to LPS (100 ng/mL) and CHX (50 μg/mL) for various times as indicated. Activation of Akt (A) and ERK (B) was determined by immunoblotting for phospho-Akt or phospho-ERK, respectively. Membranes were stripped and reprobed for α-tubulin as a loading control. (C) Wild-type or TRAF6^-/- MEFs were stimulated with LPS only (100 ng/mL), and activation of Akt was determined by immunoblotting for phospho-Akt. Membranes were stripped and probed with α-tubulin as a loading control. Results are representative of 3 independent experiments.