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Figure S1. HIP1 acts a transcriptional coregulator for other nuclear hormone receptors. To assess a role for HIP1 in ER activity, COS-7 cells were transiently transfected with the GAL4-TK luciferase reporter and either pGAL4DBD-ER{alpha} (A) or pGAL4DBD-ERß (B) together with pcDNA3-HIP1 or pCMV-ß-p300 as a positive control. Cells were incubated in steroid-depleted phenol red-free media in the presence and absence of 1 nM estradiol. After 48 h, cells were lysed and luciferase assays were performed as described for the AR. To assess a role for HIP1 in other nuclear hormone receptor function, COS7 cells were transiently transfected with increasing concentrations of pcDNA3-HIP1 together with 50 ng pcDNA3-GR and 100 ng MMTV reporter (C). After transfection, the cells were incubated in steroid-depleted media in the presence and absence of 10 nM of the GR ligand dexamethasone before luciferase analysis. (D) HIP1 does not activate the ligand-binding domain (LBD) of the GR upon the GAL4 reporter. Transient transfections were performed as before, but including 50 ng of the GAL4-DBD-GR-LBD fusion and 100 ng of the GAL4-DBD-responsive TKLuc reporter. Data represent the means of three independent experiments ± SD per panel/set of conditions.