Representative 3D aMT length tracking results, d istribution of Kip2 mutants along microtubules, and SDS-PAGE of proteins used in in vitro assays. (A and B) Representative 3D aMT length was extracted from control (A) and kip2Δ (B) cells using Bik1-3xsfGFP and Spc72-eGFP as microtubule plus- and minus-end markers, respectively. The detection limit (666.7 nm due to the microscope resolution) is marked as a magenta dashed line. Rescue (green) and catastrophe (black) events are marked with triangles. The lifetime of each aMT is indicated with a black line with two arrowheads. (C) Representative images of full-length Kip2-3xsfGFP and Kip2-∆motor-3xsfGFP accumulation on microtubule tips. (D) The chimera protein is composed of Kip3-MD and Kip2-NMD, then C-terminally fused with mNeonGreen, expressed from KIP3 locus. The red arrowhead marks the plus-end of the aMT. SPBs are visualized with Spc72-GFP. Corresponds to Video 1. The time-lapse movie has 1.07 s intervals; only representative time points are shown here. Scale bar, 2 µm. (E) Coomassie-Blue-stained SDS-PAGE analysis of indicated proteins. Details of the constructs are summarized in Table S3. The Kip2-NMD image was cropped from the gel shown in Fig. 2 C. Source data are available for this figure: SourceData FS1.