FIG 5.

Expression of hepatocyte-specific genes is reduced in DGAT1-silenced cell lines. (A) HNF4α mRNA levels in DGAT1-silenced cell lines were determined by real-time quantitative PCR (n = 3) (upper graph), and HNF4α protein expression was assessed by immunoblotting (lower blots). (B) mRNA expression of representative hepatic differentiation markers in control and DGAT1-silenced cell lines. Data are standardized to β-actin mRNA levels (n = 3). Bar graphs represent means ± SEM. *, P < 0.001 compared to the control. (C) Immunoblot analysis was performed to evaluate HNF4α expression in control and DGAT2 knockdown Huh-7.5 cell lines after 40 days of lentivirus shRNA transduction. Data are representative of two independent experiments. (D) Huh-7.5 cells were transduced with lentivirus harboring shRNA-DGAT1-1402, and temporal changes in protein expression were examined during the establishment of a stable cell line. The cell pellets were harvested at days 10, 25, 32, and 45 after transduction and subjected to immunoblot analyses to detect DGAT1, HNF4α, and CLDN1. Data are representative of two independent experiments.