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. 1999 Oct 12;96(21):11866–11871. doi: 10.1073/pnas.96.21.11866

Figure 7.

Figure 7

Rb interacts with BRCA1 protein. (A and C) Pull-down assay with the use of recombinant GST–Rb or GST–BRCA1 polypeptides. GST fusion proteins were incubated with U2OS (A) or SK-N-SH (C) cell lysates, and bound BRCA1 or Rb proteins were identified by Western blot (WB). Protein (100 μg) was loaded into WCE lanes. Approximately 1 mg of protein was used for each pull-down reaction. In A, GST-Grb7 protein was used as a negative control; in the last lane, the GST–RbABC/BRCA1 protein complex was released from the glutathione–Sepharose with glutathione (GSH) before loading as described in Methods. (B and D) Coimmunoprecipitation of pRb and BRCA1. Rb or BRCA1 proteins were immunoprecipitated from U2OS cell lysate by using anti-Rb (C-36 or C-15) (B) or anti-BRCA1 (MS110) (D). Normal mouse or rabbit IgG or anti-CD54 antibody was used as a negative control. Total protein (100 μg) was loaded into WCE lanes and ≈ 2 mg of protein was used for each immunoprecipitation.