Overexpression of murine RORα4 and γ1 dramatically increases
endogenous CYP2C8 mRNA in HepG2 cells. Adenoviruses containing
FLAG-mRORα4 and the control LacZ (A) or FLAG-mRORγ1 and the
control GFP/β-Gal (B) were utilized to infect HepG2 cells in triplicate
for 1 to 4 days with a dose of 1000 particles/cell. Cells were harvested and
used to isolate total RNA. cDNAs were synthesized with MMLV, and then
real-time quantitative PCR analyses were performed to determine the expression
of CYP2C8 and TBP. Endogenous CYP2C8 mRNA gradually
increased after infection of HepG2 cells with adenoviruses containing ROR over
those infected with LacZ significantly (**, p < 0.01;
***, p < 0.001). Data represent means ± S.E.
(n = 9). All data are normalized to TBP levels. The same ROR viruses
and the GFP control were used to infect HepG2 cells in six-well plate in
duplicates. After 2 days, the whole-cell lysates were prepared, and the
Western blot was performed to detect the expression of exogenous FLAG-tagged
ROR protein (C and D).