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. 2014 Apr 4;3(5):326–331. doi: 10.1242/bio.20147955

Fig. 4. Mutation of the clathrin-box site is sufficient to block interaction with clathrin-box motif-containing proteins.

Fig. 4.

Results of a typical binding experiment to analyse the role of the four interaction sites on the NTD of CHC. Wild-type MBP-CHC(1–1074)-His6 or indicated mutants were incubated with (A) GST or GST-β2 adaptin (616–951) or (B) GST or GST-TACC3-His6 that were phosphorylated with Aurora A/TPX2, before pull-down on glutathione beads. Bound, and a sample of unbound, material was separated by SDS-PAGE and transferred to nitrocellulose for western blotting using an anti-CHC (TD.1) antibody. Markers indicate position of 150 kDa.