Figure 1.

Interaction between EHD1 and SNX17. A, HeLa cell lysates were incubated at 4 °C overnight with either anti-SNX17, anti-EHD1, or anti-EB3 antibodies (from left to right). Protein G beads were then added to the lysate–antibody mixture at 4 °C for 4 h. Bound proteins were then eluted by boiling at 95 °C in β-mercaptoethanol–containing loading buffer, separated by SDS-PAGE, and immunoblotted with anti-SNX17 antibodies (left panel), anti-EHD1 antibodies (center panel), or anti-EB3 antibodies (right panel). Input lysates (20%) are depicted on the left of the immunoblots. h.c., immunoglobulin heavy chain. B, purified His-EHD1 was bound to Ni²⁺-NTA beads prior to incubation with either GST alone or GST-SNX17. Bound proteins were then eluted by boiling at 95 °C in β-mercaptoethanol–containing loading buffer, separated by SDS-PAGE, and immunoblotted with anti-His (left panel) or anti-GST antibodies (center and right panels). Input refers to the amounts of purified GST and GST-SNX17 used for incubation with His-EHD1. Data shown are representative of three independent experiments.