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. 2009 Apr;23(4):466–474. doi: 10.1210/me.2008-0295

Fig. 5.

Fig. 5.

BARD1/BRCA1 induces ubiquitination of LXRα. A, LXRα-expressing HEK293 cells were successively transfected with BARD1/BRCA1 and then treated with cycloheximide for the indicated periods. B, Transfected HEK293 cells with LXRα- and HA-tagged ubiquitin expression vectors were sequentially transfected BRCA1 and/or BARD1. After treatment of T0901317 (5 μm) and MG132 (20 μm) for 3 h, total cell lysates were subjected to in vivo ubiquitination assays. C, LXRα-expressing HEK293 cells were transfected with siRNA duplex for negative control (siControl) and BARD1 (siBARD1), and the stability of LXRα protein was monitored by Western blotting. D, The siControl and siBARD1 siRNA duplexes were transfected into HEK293 cells expressing LXRα and ubiquitin, after which in vivo ubiquitination assay was performed. CHX, Cycloheximide; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IB, immunoblotting; IP, immunoprecipitation.