Figure S3.
Targeting MT motors or vesicle formation affects THN migration (related to Figs. 4 and 5).(A) Endogenous MT acetylation is low in 29-hpf embryos but can strongly be increased by overexpression of aTat1. Scale bar, 10 µm. (B) Quantification of MT acetylation in citrine- or aTat1-expressing THNs. Percentage of MT acetylation–positive cells out of all THNs expressing the genetic marker per embryo is shown. n = 7 embryos (citrine), n = 9 embryos (aTat1). (C) When dynein is inactivated, leading processes are still able to grow along the MHB. Box indicates magnified region on the right. Scale bars, 20 µm. (D) The overexpression of a dominant-negative fragment of Lis1a decreases THN motility. Arrows and arrowheads indicate the start and end points on the right. Scale bars, 20 µm. (E) The reduction in THN motility after quantification is less pronounced than in dynein inactivation by p50. n = 24 embryos/145 tracks for control, n = 14 embryos/191 tracks for Lis1N-expressing embryos, P = 0.027. (F) Brefeldin A treatment disassembles the Golgi apparatus very efficiently within 2 h, while Dynasore blocks endosome formation immediately after injection (G). Blue boxes indicate regions magnified on the right. Scale bars represent 20 µm in the overview and 10 µm in the magnified region. (H) Overexpression of wild-type kinesin-3 Kif1C does not impair THN migration, but the rigor T106N mutation reduces motility. Scale bars, 20 µm. (I) Similarly, the overexpression of WT Rab11a does not interfere with THN motility, but the dominant-negative S25N mutation reduces it. Scale bars, 20 µm. Elapsed time in minutes is given at the top of the respective images. MHB, solid line; URL, dotted line. Colored arrows on the right indicate the start point of an individual THN, and arrowheads of the same color indicate the end point after 180 min. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05.

Targeting MT motors or vesicle formation affects THN migration (related to Figs. 4 and 5 ). (A) Endogenous MT acetylation is low in 29-hpf embryos but can strongly be increased by overexpression of aTat1. Scale bar, 10 µm. (B) Quantification of MT acetylation in citrine- or aTat1-expressing THNs. Percentage of MT acetylation–positive cells out of all THNs expressing the genetic marker per embryo is shown. n = 7 embryos (citrine), n = 9 embryos (aTat1). (C) When dynein is inactivated, leading processes are still able to grow along the MHB. Box indicates magnified region on the right. Scale bars, 20 µm. (D) The overexpression of a dominant-negative fragment of Lis1a decreases THN motility. Arrows and arrowheads indicate the start and end points on the right. Scale bars, 20 µm. (E) The reduction in THN motility after quantification is less pronounced than in dynein inactivation by p50. n = 24 embryos/145 tracks for control, n = 14 embryos/191 tracks for Lis1N-expressing embryos, P = 0.027. (F) Brefeldin A treatment disassembles the Golgi apparatus very efficiently within 2 h, while Dynasore blocks endosome formation immediately after injection (G). Blue boxes indicate regions magnified on the right. Scale bars represent 20 µm in the overview and 10 µm in the magnified region. (H) Overexpression of wild-type kinesin-3 Kif1C does not impair THN migration, but the rigor T106N mutation reduces motility. Scale bars, 20 µm. (I) Similarly, the overexpression of WT Rab11a does not interfere with THN motility, but the dominant-negative S25N mutation reduces it. Scale bars, 20 µm. Elapsed time in minutes is given at the top of the respective images. MHB, solid line; URL, dotted line. Colored arrows on the right indicate the start point of an individual THN, and arrowheads of the same color indicate the end point after 180 min. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05.

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