Figure 6. Inhibition of AKT and ERK signaling pathways and transcriptional activity of NF-κB by TSC-22.

(A) HeLa cells were transiently transfected with pcDNA4-TSC-22, then harvested at the indicated time points and analyzed by western blot. The expression levels of pERK, ERK, Elk-1, Egr-1, pAKT, AKT, pGSK-3β, and GAPDH were measured. (B) HeLa cells were transiently transfected with TSC-22 in a dose-dependent manner. The activity of the NF-κB promoter was measured using a luciferase reporter gene assay. The firefly luciferase and Renilla luciferase values were determined. Firefly luciferase levels were normalized to Renilla luciferase levels. (C) TSC-22-overexpressing cells were harvested at 24-hour intervals and NF-κB protein levels were evaluated using western blotting.