Fig. 7. Mapping of the centaurin-₁ and KIF13B binding sites using a GST pull-down assay. (A) Lysates of COS cells expressing GFP or GFP-tagged centaurin-₁ (cent₁) or its deletion mutants were incubated with either GST or GST-KIF13B coupled to glutathione beads. (B) The lysates of COS cells expressing GFP-tagged KIF13B or KIF13B were incubated with either GST or GST-centaurin-₁ coupled to glutathione-beads. After washing the beads, the bound proteins were separated on SDS-PAGE, blotted onto PVDF membranes, and probed with an anti-GFP antibody. 5% of the inputs (cell lysates) are also shown for each of the sets of experiments. These experiments were repeated three times with similar results.