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. 2009 Nov 15;20(22):4686–4695. doi: 10.1091/mbc.E08-11-1123

Figure 2.

Figure 2.

Hice1 binds to the Hec1/Nuf2 complex. (A and B) Coomassie Blue staining of various purified His-tagged Hice1 versions and GST-Nuf2 (A), and Hec1/GST-Nuf2 dimer (B). Hice1ΔCoil1, amino acid 150-228 deleted; Hice1ΔCoil2, amino acids 263-329 deleted. (C) In vitro binding assay using the above purified proteins (detailed in Materials and Methods). Equal amounts of Hice1-FL, Hice1ΔCoil1, and Hice1ΔCoil2 were used to interact with Hec1/GST-Nuf2 dimer (lanes 1–3) or GST-Nuf2 (lanes 4–6). Interacting proteins were then pull downed by glutathione-Sepharose beads and subjected to SDS-PAGE followed by Western blot. (D) Hice1 coimmunoprecipitated with endogenous Nuf2 and Hec1. HeLa cell extract was subjected to coimmunoprecipitation assay with indicated antibodies. The image showed the Western blot developed with respective antibodies.