Centriole loss restrains microtubule remodeling during early axon developmentA.Figure 4A. Example stills from a spinning-disk time-lapse recording of a neurite transfected with mRFP and GFP-MT+TIP. The top panel is a still of a neurite in mRFP, showing neurite morphology. The other panels show moving GFP-MT+TIP comets pointing in either an anterograde direction (green arrowheads) or retrograde direction (blue arrowheads). P indicates the proximal direction and D the distal direction of the neurite. Timestamp in minutes:seconds given on bottom right. Scale bar = 5 µm. B. Kymographs and schematic representations of time-lapse recordings of the distal axon as shown in (A), for different time points (day 7; day 13) and conditions (control; Centrinone-B). Scale bar = 5 µm. C C. Quantifications of the percentage of neurons exhibiting uniform, or non-uniform comet orientations in the anterograde direction in the distal axon at day 7. n = 15-17 cells in 4 independent experiments. F. Quantifications of the percentage of neurons exhibiting uniform, or non-uniform comet orientations in the anterograde direction in the distal axon at day 13. n = 11-18 cells in 4 independent experiments. Data information: Data represent mean ± SEM. Chi-square-test ; ** p<0.005, * p<0.05, ns p≥0.05. D. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon at day 7. n = 15-17 cells in 4 independent experiments. G. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon at day 13. n = 11-18 cells in 4 independent experiments. Data information: Data represent mean ± SEM. unpaired t-test ; ** p<0.005, * p<0.05, ns p≥0.05. E. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon at day 7. n = 15-17 cells in 4 independent experiments. H. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon at day 13. n = 11-18 cells in 4 independent experiments. Data information: Data represent mean ± SEM. unpaired t-test ; ** p<0.005, * p<0.05, ns p≥0.05. I. Schematic representation of microtubule laser-severing (LS) experiments. J. Example stills from a spinning-disk time-lapse recording of a neurite transfected with mRFP and GFP-MT+TIP. Red line denotes the location of LS. Scale bar = 5 µm. K. Kymographs and schematic representations of time-lapse recordings of the distal axon as shown in (J) following LS, for different time points (day 7; day 13) and conditions (control; Centrinone-B). Red line and red arrowhead denote location and time of LS. Scale bar = 5 µm. L L. Quantifications of the percentage of neurons exhibiting uniform, or non-uniform comet orientations in the anterograde direction in the distal axon following LS at day 7. n = 20-22 neurons in 3 independent experiments. O. Quantifications of the percentage of neurons exhibiting uniform, or non-uniform comet orientations in the anterograde direction in the distal axon following LS at day 13. n = 24-27 neurons in 3 independent experiments. Data information: Data represent mean ± SEM. Chi-square-test ** p<0.005, * p<0.05, ns p≥0.05. M. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon following LS at day 7. n = 20-22 cells in 3 independent experiments. P. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon following LS at day 13. n = 24-27 cells in 3 independent experiments. Data information: Data represent mean ± SEM. unpaired t-test ; ** p<0.005, * p<0.05, ns p≥0.05. N. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon following LS at day 7. n = 20-22 cells in 3 independent experiments. Q. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon following LS at day 13. n = 24-27 cells in 3 independent experiments. Data information: Data represent mean ± SEM. unpaired t-test ; ** p<0.005, * p<0.05, ns p≥0.05. R. Schematic representation of the proposed orientation of microtubules in the distal axon in control conditions, and following Centrinone-B treatment.