Figure 4. Ajuba enhances LXRα target gene expression in HepG2 cells and mouse primary hepatocytes.

A, Depletion of Ajuba in HepG2 cells using lentiviral shRNA specifically targeting Ajuba. The cells were selected with puromycin for 2 weeks, and the resulting pooled cells were treated with T090 (1μM) or DMSO for 24 hours and total RNAs were extracted, which were subjected for qRT-PCR analysis for SREBP-1C and SCD-1 expression. B, Ajuba was stably expressed in HepG2 cells via viral infection followed by puromycin selection. The expression of Ajuba was verified by Western blotting, and pooled cells were analyzed as in A. C, Mouse primary hepatocytes isolated from C57BL/6 were infected with viral supernatants containing shRNA-specific targeting mouse Ajuba and subsequently selected with puromycin to eliminate noninfected cells. Ajuba protein level was examined by Western blot analysis, and pooled cells were subjected for analysis as same as above. Relative mRNA levels were normalized to β-actin expression. Experiments were repeated 3 times and results shown are averages of 3 independent experiments.