Interdependence of XMAP215^msps and Sentin localization. (A) XMAP215^msps recruits Sentin to MT plus ends and the MT lattice independent of EB1. However, GFP-Sentin localization was far weaker in the absence of EB1. Inset shows enlarged view of the boxed area. Arrows in the inset indicate GFP-Sentin signals. (B) Coprecipitation of XMAP215^msps and Sentin. XMAP215^msps-HA and GFP-Sentin were coexpressed in Sf21 cells, and immunoprecipitation (IP) was performed with anti-HA antibodies. Protein bands after Coomassie staining are displayed (arrow, Sentin; sphere, XMAP215^msps). (C) Some shrinking MTs showed EB1-GFP signals in the presence of XMAP215^msps and Sentin. (D) Sentin facilitates localization of a low concentration of XMAP215^msps (5 nM) to the plus ends. (right) Quantification of the XMAP215^msps-GFP intensity at the growing MT tip in the presence or absence of Sentin. The data shown are from a single representative experiment out of two repeats. A.U., arbitrary unit. Bars: (horizontal) 5 µm; (vertical) 1 min.