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. 2001 Feb 1;15(3):328–339. doi: 10.1101/gad.185901

Figure 5.

Figure 5

Interaction partners for the WT1 17AA region. (A) Diagram of GST-WT1 fusion proteins (residues 245–297) either lacking (GST D−) or containing (GST D+) the 17AA motif. (B) Protein affinity chromatography of HeLa nuclear extract fractionated over a GST, GST D−, or GST D+ column. After extensive washing, the bound fraction was eluted and analyzed by immunoblottting with either anti-par4 (top) or anti-TBP (bottom) antibodies. (C) HeLa nuclear extract was fractionated over either a GST D− or a GSD D+ column, and after a brief low-salt wash (250 mM KCl) and a subsequent wash (1 M KCl), the presence of par4 was assessed in each fraction by immunoblotting with anti-par4 antibodies. (D) A GST pull-down assay was performed with immobilized GST-par4 and a bacterial lysate containing the recombinant GAL4 derivatives indicated. The bound fraction was analyzed by immunoblotting with anti-GAL4 antibodies. I is 10% of the input into each pull-down assay.