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. 2001 Aug 7;98(17):9587–9592. doi: 10.1073/pnas.171174298

Figure 2.

Figure 2

Ectopic expression of wild-type BRCA1 in Brca1-deficient MEFs restores ligand-independent repression of ERα transactivation in a histone deacetylase (HDAC)-dependent manner. (A and B) Brca1−/− MEFs in estrogen-free media were transfected with pERE-TK-Luc without (−) or with (+) pRSV-ERα, pCDNA3.1-BRCA1 expressing wild-type human BRCA1 (WT), or pCDNA3.1-BRCA1 derivatives bearing missense mutants A1708E, Q356R, A1708E/Q356R, or C64G before assay for luciferase activity. Where indicated, trichostatin A (TSA; 330 nM) was also included. (C) Brca1−/− MEFs in estrogen-free media were untransfected (lane 1) or cotransfected with expression vectors for ERα and either wild-type BRCA1 (lane 2) or various BRCA1 mutant derivatives (lanes 3–6) as indicated. Cells were lysed, and immunoprecipitated BRCA1 and ERα were subjected to immunoblot analysis using antibodies specific for BRCA1 (Top) or ERα (Middle). Immunoblot analysis of the nuclear matrix protein p84 (Bottom) indicates that nearly equivalent amounts of each cell lysate were used in the immunoprecipitations.