Figure S1.
Map4MTB labels MTs in zebrafish (related to Fig. 2).(A) Schematic representation of full-length Map4 and Map4MTB fragment. (B and C) In THNs, Map4MTB decorates MTs, which extend from the rear of the cells. Colored arrows indicate bundles in phase 2, arrowheads indicate bundles in phase 1 THNs, and dots mark the corresponding nuclei. Box indicates the region magnified in C. Elapsed time in minutes. Scale bars represent 20 µm in B and 10 µm in C. See also Video 1. (D) THNs coexpressing EB3-GFP and Map4MTB-mKate2 show colocalization in a mitotic spindle. Scale bar, 2 µm. (E) Map4MTB-labeled structures are capped with EB3. Box in overview image shows the position of the magnified region on the right. In the image column on the right, arrowheads point to a growing MT tip. Elapsed time in seconds. Scale bars represent 10 µm in the overview and 2 µm in magnified images. (F) MTOC position (arrow) can be inferred by Map4MTB-mediated MT labeling. Scale bar, 2 µm. (G) Examples of MT growth in hindbrain neurons expressing EB3-GFP, Map4MTB-mScarlet, or both. Arrowheads in images on the left indicate MT tips shown in frame montages on the right. Dotted lines indicate growth of MT tips. Elapsed time in seconds. Scale bars represent 5 µm in overview images and 2 µm in montages. (H) Kymographs of the examples shown in G. Horizontal scale bars, 1 µm; vertical bars, 10 s. (I and J) MT plus ends in THNs expressing EB3-GFP appear to grow two times faster than in Map4MTB-mScarlet overexpressing THNs using 2D tracking (I) or kymograph analysis (J). Mean growth speeds per cell are shown. Box represents 25–75% of all values, the whiskers 1.5 times the quartile. Median is shown as horizontal bar, mean as square box. 2D tracking (I): N (EB3) = 12, mean = 0.116 µm/s, N (Map4MTB) = 14, mean = 0.086 µm/s, P = 0.027 to EB3, F = 5.575; N (both) = 12, mean = 0.124 µm/s, P = 0.124, F = 2.299 to Map4MTB, P = 0.774, F = 0.084 to EB3. Kymograph analysis (J): N (EB3) = 12, mean = 0.158 µm/s, N (Map4MTB) = 14, mean = 0.096 µm/s, P = 0.008 to EB3, F = 8.355; N (both) = 12, P = 0.938, F = 0.006 to Map4MTB, mean = 0.097 µm/s, P = 0.008, F 8.577 to EB3. Significance level in one-way ANOVA: *, P < 0.05; **, P < 0.01; ns, not significant.

Map4MTB labels MTs in zebrafish (related to Fig. 2 ). (A) Schematic representation of full-length Map4 and Map4MTB fragment. (B and C) In THNs, Map4MTB decorates MTs, which extend from the rear of the cells. Colored arrows indicate bundles in phase 2, arrowheads indicate bundles in phase 1 THNs, and dots mark the corresponding nuclei. Box indicates the region magnified in C. Elapsed time in minutes. Scale bars represent 20 µm in B and 10 µm in C. See also Video 1. (D) THNs coexpressing EB3-GFP and Map4MTB-mKate2 show colocalization in a mitotic spindle. Scale bar, 2 µm. (E) Map4MTB-labeled structures are capped with EB3. Box in overview image shows the position of the magnified region on the right. In the image column on the right, arrowheads point to a growing MT tip. Elapsed time in seconds. Scale bars represent 10 µm in the overview and 2 µm in magnified images. (F) MTOC position (arrow) can be inferred by Map4MTB-mediated MT labeling. Scale bar, 2 µm. (G) Examples of MT growth in hindbrain neurons expressing EB3-GFP, Map4MTB-mScarlet, or both. Arrowheads in images on the left indicate MT tips shown in frame montages on the right. Dotted lines indicate growth of MT tips. Elapsed time in seconds. Scale bars represent 5 µm in overview images and 2 µm in montages. (H) Kymographs of the examples shown in G. Horizontal scale bars, 1 µm; vertical bars, 10 s. (I and J) MT plus ends in THNs expressing EB3-GFP appear to grow two times faster than in Map4MTB-mScarlet overexpressing THNs using 2D tracking (I) or kymograph analysis (J). Mean growth speeds per cell are shown. Box represents 25–75% of all values, the whiskers 1.5 times the quartile. Median is shown as horizontal bar, mean as square box. 2D tracking (I): N (EB3) = 12, mean = 0.116 µm/s, N (Map4MTB) = 14, mean = 0.086 µm/s, P = 0.027 to EB3, F = 5.575; N (both) = 12, mean = 0.124 µm/s, P = 0.124, F = 2.299 to Map4MTB, P = 0.774, F = 0.084 to EB3. Kymograph analysis (J): N (EB3) = 12, mean = 0.158 µm/s, N (Map4MTB) = 14, mean = 0.096 µm/s, P = 0.008 to EB3, F = 8.355; N (both) = 12, P = 0.938, F = 0.006 to Map4MTB, mean = 0.097 µm/s, P = 0.008, F 8.577 to EB3. Significance level in one-way ANOVA: *, P < 0.05; **, P < 0.01; ns, not significant.

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