Figure 2.

Analysis of the regions in BRCA1 and EZH2 responsible for their interaction. (A) Schematic diagram of the five GST-BRCA1 fusion proteins (BR1 to BR5). NLS, nuclear localization signal. (B) GST pull-down assay detecting the regions of BRCA1 that bind to PRC2 proteins. Top, western blot analysis of in vitro transcribed and translated Polycomb proteins EZH2, SUZ12, EED and Ezh1 pulled down by GST or GST-BRCA1 fusion proteins BR1-BR5. ^#Nonspecific proteins reacted with anti-EZH2 antibody. Bottom, Coomassie blue staining of GST and GST-BRCA1 fusion proteins BR1–BR5. Asterisks indicate proteins with correct molecular masses. (C) Top, western blot analysis of Ezh2 and EED in baculovirally expressed PRC2 complex pulled down by GST or GST-BRCA1 fusion proteins. Bottom, Coomassie blue staining of GST and GST-BRCA1 fusion proteins BR1–BR5. Asterisks indicate proteins with correct molecular masses. (D) GST pull-down assay detecting the regions of EZH2 that bind to BRCA1. Top, schematic diagram of the four GST-EZH2 fusion proteins designated as EZ1 to EZ4. EBD, EED binding domain; ncRBD1, ncRNA binding domain 1. Middle, western blot analysis of baculovirally expressed BRCA1 pulled down by GST or GST-EZH2 fusion proteins EZ1-EZ4. ^#Nonspecific proteins reacted with anti-BRCA1 antibody. Bottom, Coomassie blue staining of GST and GST-EZH2 fusion proteins EZ1–EZ4. Asterisks indicate proteins with correct molecular mass.

Source data for this figure is available on the online supplementary information page.

Source Data for Figure 2 ^{(2.5MB, tif)}