Fig. 4.

Inhibition of TNF-α activity by HDAC knockdown or chemical inhibitor. A, HDAC3 protein in knockdown cells. HDAC3 protein was determined in whole-cell lysates of HEK 293 cells by Western blot 48 h after transfection with the HDAC3 RNAi plasmid. B, Restoration of Pepck promoter by HDAC3 knockdown. The assay was conducted in 293 cells. C, Restoration of Pepck mRNA by TSA. mRNA was determined using qRT-PCR after 4 h of treatment. D, Restoration of PEPCK protein expression by TSA. The mature 3T3-L1 cells were pretreated with TSA (50 nm) before troglitazone and TNF-α treatment. PEPCK protein was examined in whole-cell lysates by Western blot. The PEPCK protein signal was quantified and presented in the bar graph. In this figure each bar represents mean ± sem (n = 3).