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. Author manuscript; available in PMC: 2007 Mar 12.
Published in final edited form as: Mol Genet Metab. 2006 Sep 7;90(1):97–111. doi: 10.1016/j.ymgme.2006.07.011

Fig. 5.

Fig. 5

TGIF HPE mutants interact with RXRα and affect repression of retinoic acid regulated transcription. (A) COS1 cells were transfected with T7-RXRα and Flag-TGIF wildtype or HPE mutant construct, as indicated. Protein complexes in lysates were isolated on anti-Flag agarose and analyzed by western blot for the presence of co-precipitating RXRα. Coprecipitating RXR is indicated with an arrow, the Ig heavy chain with a bar. A portion of the lysates was subjected to direct western blot analysis to monitor protein expression (below). (B and C) HepG2 cells were transfected with a DR1-TATA-luc luciferase reporter and RXRα (control and experimental), together with the indicated TGIF wildtype or HPE mutant expression constructs in the experimental lanes. Cells were either left untreated (black bars) or treated with 9-cis-retinoic acid (9C-RA) for 24 hours prior to analysis (striped bars). Reporter luciferase activity (normalized to Renilla luciferase activity) was assayed 40 hours after transfection and is presented, in arbitrary units, as the mean +/− s.d., of duplicate transfections.