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. 2011 Oct 3;195(1):71–86. doi: 10.1083/jcb.201011112

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© 2011 Willenborg et al.

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Figure 6. — FIP5 binds to SNX18-LC domain. (A and B) The affinity of FIP5 and SNX18 binding as determined by ITC. N.D., not detected. (C) Glutathione beads were coated with GST-SNX18, GST-SNX9, or GST alone and incubated with 6His-FIP5 or 6His-FIP3 in the presence or absence of a fivefold excess of Rab11a-GTP. The amount of bound 6His-FIP3 or 6His-FIP5 was determined by immunoblotting with anti-FIP3 or anti-FIP5 antibodies. (D) Glutathione beads were coated with GST-SNX18 or GST alone and incubated with 6His-FIP5 in the presence of increasing concentrations of soluble recombinant SNX9. The amount of bound 6His-FIP5 was determined by immunoblotting with anti-FIP5 antibodies. (E) Schematic representation of the FIP5-binding domain in SNX18 as determined by glutathione bead pull-down assays.