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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: Traffic. 2016 Jul 24;17(10):1091–1109. doi: 10.1111/tra.12423

Figure 2. The PA-binding domain DEP reduces Sec18p binding to membranes.

Figure 2

(A) Liposomes of the indicated compositions were incubated without Dvl2-DEP or with 20 µM Dvl2-DEP for 5 min at 30°C. Recombinant His6-Sec18p (2 µg) was added to the sample and incubated for 10 min at 30°C. (B) GST-DEP titration and Sec18p binding to liposomes was performed as described above in the presence of the indicated concentrations of GST-DEP. (C) Vacuoles were harvested from wild type DKY6281 and incubated with GST-DEP, GST-C1b domain, or GST-ENTH domain. Bound and unbound fractions were separated by centrifugation and resolved by SDS PAGE. Bar graphs show the percentage of unbound Sec18p in each sample normalized to wild type control. * P<0.05; ** P<0.001; *** P<0.0001.