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. 2009 Nov 6;285(1):595–607. doi: 10.1074/jbc.M109.042697

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FIGURE 5. — ASH2L and DPY-30 interact through their SDI and Dpy-30 domains. A, schematic representation of ASH2L and DPY-30 and corresponding mutants used. Positions of annotated domains are depicted. B, GST fusion binding assays were performed to determine critical regions for ASH2L and DPY-30 interaction. Purified full-length GST-DPY-30, GST-Dpy-30 Δ77–80, or GST-Dpy-30 L65E,L66E was used to pull down bacterially expressed His₆-ASH2L or His₆-Ash2L mutants from whole cell extracts. GST and glutathione-agarose beads (Agarose) were used as controls for nonspecific binding. Input and bound fractions of His₆-ASH2L and His₆-Ash2L mutants were detected by immunoblotting using anti-His monoclonal antibody. Immunoblots were stained with Coomassie Blue to indicate the amount of GST, GST-DPY-30, GST-Dpy-30 Δ77–80, and GST-Dpy-30 L65E,L66E proteins used.