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. 2006 Oct 27;400(Pt 1):53–62. doi: 10.1042/BJ20060774

Figure 2. Determination of regions within hMYH involved in hHus1 binding.

Figure 2

(A) The GST pull-down assay was employed using various GST–hMYH constructs to determine the binding regions within hMYH for interaction with hHus1 in HeLa nuclear extracts (0.4 mg). Lane 1 contains 15% of input (60 μg) of nuclear extracts (NE). In lanes 2–7, GST fusion proteins, indicated on top of each lane, were used in the pull-down experiments. INT stands for intact hMYH. Western-blot analyses of the pellets were performed with the antibody against hHus1. A control was run concurrently with immobilized GST alone (lanes 8). The results represent four separate experiments in which 15% of total input is used as a standard and GST alone is used as a negative control for each pull-down analysis. (B) Graphic depiction of GST–hMYH constructs and the hHus1 binding to the hMYH fusion proteins. The amino acid residues of hMYH in the GST constructs are indicated. The ‘+’ and ‘–’ listed on the right of each construct indicate the presence and absence of hHus1 in the pellets (PPT) of GST beads respectively.