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. 2007 Oct 22;28(1):140–153. doi: 10.1128/MCB.00662-07

FIG. 1.

FIG. 1.

Hbo1, ING4, ING5, and p53 associate in shared protein complexes in cells. (A) Hbo1 and p53 specifically coprecipitate from MCF7 whole-cell extracts. (Left) Results of Western blotting using a monoclonal anti-p53 antibody (α-p53) to detect the presence of p53 in the labeled samples. Lane 1, 13% of the input MCF7 whole-cell extract used for the other immunoprecipitations; lane 2, control immunoprecipitation (IP) prepared using purified rabbit IgG; lane 3, precipitates prepared with anti-Hbo1 antibody alone; lane 4, Hbo1 antibody plus the specific antigenic peptide L74-1; lane 5, Hbo1 antibody plus a nonspecific peptide, MBP4-14; lane 6, Hbo1 antibody plus ethidium bromide (EtBr). (Right) Results of Western blotting using anti-Hbo1 antibody to detect the presence of Hbo1 in the labeled samples. Lanes 7, 10, and 13, MCF7 cells that were mock transfected; lanes 8, 11, and 14, cells transfected with FLAG vector without insert; lanes 9, 12, and 15, cells transfected with vector expressing FLAG-tagged Hbo1; lanes 7 to 9, samples of 4% of the input extracts used for the other immunoprecipitations; lanes 10 to 12, immunoprecipitates prepared with anti-p53 antibody (PAb 1801) (α-p53); lanes 13 to 15, immunoprecipitates prepared with anti-PMS2 antibody of the same IgG1 subtype as PAb 1801 (α-PMS2). (B) Hbo1 and ING4 coprecipitate, and Hbo1, ING5, and p53 coprecipitate. (Left) Results of Western blotting of anti-Hbo1 immunoprecipitations using anti-ING4 and anti-ING5 antibodies (lanes 1 to 3). Lane 1, whole-cell extract (WCE) which was first immunoprecipitated with anti-ING4 antibody (top row) or anti-ING5 antibody (bottom row) and then run together with the control (lane 2) and anti-Hbo1 immunoprecipitates (lane 3) for Western blotting; lane 2, immunoprecipitates prepared with control preimmune serum; lane 3, immunoprecipitates prepared with anti-Hbo1 antibody. (Right) Results of Western blotting using anti-Hbo1 and anti-p53 antibodies (lanes 4 to 7). Lane 4, 1% of the whole-cell extract; lane 5, immunoprecipitates prepared with control goat IgG; lane 6, immunoprecipitates prepared with anti-ING4 antibody; lane 7, immunoprecipitates prepared with anti-ING5 antibody.

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