Fig. 4.

CITCO promotes the nuclear translocation and target gene interaction of hCAR1+A in immortalized cells. COS1 were transfected with 1 μg of EYFP-hCAR1, EYFP-hCAR3, or EYFP-(hCAR1+A) as outlined under Materials and Methods. Transfected cells were then treated with either 0.1% DMSO or 1 μM CITCO for 24 h. A, confocal images illustrate representative localization and translocation of different EYFP-hCAR expression after vehicle control or CITCO treatment. B, one hundred EYFP-hCAR-expressing cells from each group were classified into cytoplasmic, nuclear, or mixed (cytoplasmic + nuclear) distributions. C, nuclear proteins (30 μg) extracted from hCAR1, hCAR3, or hCAR1+A expression vector transfected COS1 cells were subjected to hCAR immunoblot analysis. D, in a separate experiment, HepG2 cells were transfected with hCAR1 or hCAR1+A expression vector for 24 h and subsequently treated with CT (0.1% DMSO) or CITCO (1 μM) for 2 h. Harvested cells were subjected to CHIP assays as described under Materials and Methods.