Feline Lindhout

3 Centrosome-mediated microtubule remodeling during axon formation in human iPSC-derived neurons 107 Supplementary Figure 4. Centriole loss restrains microtubule remodeling during early axon development A. Kymographs and schematic representations of time-lapse recordings of dendrites, for different time points (day 7; day 13) and conditions (control; Centrinone-B). Scale bars, 5 µm. B. 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 bars, 5 µm. C. Quantifications of the number of comets per minute pointing in the anterograde direction in the proximal axon. N= 3, n= 7-11 cells. D. Quantifications of the number of comets per minute pointing in the retrograde direction in the proximal axon. N= 3, n= 7-11 cells. E. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon. N= 3, n= 8-12 cells. F. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon. N= 3, n= 8-12 cells. G. Quantifications of the ratios of comets pointing in anterograde (green) or retrograde (blue) direction in the dendrite. N=3, n=7-10 cells. H. Quantifications of the number of comets per minute pointing in the anterograde direction in the dendrite. N=3, n=7-10 cells. I. Quantifications of the number of comets per minute pointing in the retrograde direction in the dendrite. N=3, n=7-10 cells. J-L . Quantifications of the growth speed of comets in the dendrite ( J ), proximal axon ( K ), and distal axon ( L ). N=3 , n=9-2406 traces in 7-10 cells. M-O. Quantifications of the distance of run length of comets in the dendrite ( M ), proximal axon ( N ), and distal axon ( O ). N=3 , n=n=9-2406 traces in 7-10 cells. P. Kymographs and schematic representations of time-lapse recordings of laser severing (LS) experiments of dendrites, 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 bars, 5 µm. Q. 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 bars, 5 µm. R. Quantifications of the number of comets per minute pointing in the anterograde direction in the proximal axon following LS. N=3 , n=19-30 cells. S. Quantifications of the number of comets per minute pointing in the retrograde direction in the proximal axon following LS. N=3 , n=19-30 cells. T. Quantifications of the number of comets per minute pointing in the anterograde direction in the distal axon. N=3 , n=19-30 cells. U. Quantifications of the number of comets per minute pointing in the retrograde direction in the distal axon. N=3 , n=19-30 cells. V. Quantifications of the ratios of comets pointing in anterograde (green) or retrograde (blue) direction in the dendrite following LS. N=3, n=19-30 cells. W. Quantifications of the number of comets per minute pointing in the anterograde direction in the dendrite following LS. N=3, n=19-30 cells. X. Quantifications of the number of comets per minute pointing in the retrograde direction in the dendrite following LS. N=3, n=19-30 cells. Data information: Data represent mean ± SEM. One-way ANOVA including Sidak’s post-hoc analysis ( C - F , H - O , R - U , W , X ); *** p<0.001, ** p<0.005, * p<0.05, ns p≥0.05