Fig. 2. Ethanol increases TBP expression in vivo and in vitro.

(A) Ethanol increased TBP expression in HepG2-ADH cells. HepG2-ADH cells were starved in DMEM for 3 h and treated with 50mM ethanol for 1 h. The total RNA and cell lysates from these cells were extracted to determine TBP mRNA by RT-qPCT. Resultant protein lysates were subjected to perform immunoblot analysis. The antibodies were indicated (A lower panel). (B) Chronic alcohol administration in mice induced TBP expression. C57BL/6 transgenic mice harboring the HCV NS5A gene that is selectively expressed in hepatocytes were chronically fed with control diet or 3.5% of ethanol in the Lieber-DeCarli liquid diet for 12 months. Liver tissues were harvested from these mice, Cell lysates and RNA were extracted from non-tumor or tumor portions of the livers. RT-qPCR was used to measure the amounts of TBP transcript relative to GAPDH. The values represent ± SE from three independent experiments. The lysates of the livers were subjected to immunoblot analysis with antibodies of TBP and β-actin. Each group of mouse includes at least three mice. The bars represent Mean ± SE of at least three independent determinations. **: p<0.01. The fold change was calculated in each group by normalizing to mice fed with control diet.