Fig. 1: Nuclear mobility is impaired in DCX-KO cortical neurons.

A DCX contains two microtubule binding domains, DC1 and DC2. DCX-KO cells were obtained by indels introduced in Dcx’s gene sequence before DC1. CTRL0 and DCXKO1 amino acid sequences are shown from residue 41–60. The asterisk shows the indel location and the resulting amino acid sequence in DCXKO1 (cyan). B Immunoblots representative of 3 experiments, showing CTRL0 and DCXKO1 neuron lysates at 3 days of differentiation (DIV3), stained with antibodies against β3-Tubulin (Tubb3) and DCX. C CTRL0 and DCXKO1 neurons representative of 4 experiments stained for Tubb3 and DCX at DIV3. D (top panel) Still images of live neurons with a nuclear dye. Nuclei detected by Trackmate are circled (light blue), and their trajectories over the 8 h period of imaging are shown (gray). (Bottom panel) Position plots showing a sample of trajectories extracted from CTRL0 and DCXKO1 nuclei tracking. The first timepoint of all tracks is (0,0). Tracks are color-coded according to the displacement of the nucleus. Distribution of displacement for both genotypes are shown on the right end side of the panel. E Schematics showing the displacement, distance traveled, and directionality index parameters. F Distribution of mean speed achieved by CTRL and DCXKO nuclei during the imaging period. G Distribution of directionality index or efficiency of the trajectory for CTRL and DCXKO nuclei. Higher directionality indexes indicate more unidirectional movements. H Distribution of distance traveled by CTRL and DCXKO nuclei during the imaging period. For (F–H) Violin plots are split between the two control (CTRL0, light gray, and CTRL1, dark gray) and two DCXKO (DCXKO1, cyan, and DCXKO4, dark cyan) lines used in this study. These three plots were cut-off to better show the main population, but the maximum values for distance traveled, mean speed, and directionality index are 497 µm, 2.7 µm/min and 1 respectively. n = 21826 tracks for CTRL0, 24801 tracks for CTRL1, 6461 tracks for DCXKO1 and 14,837 tracks for DCXKO4. Each line was used in 3 experiments, tracks with less than 30 min of imaging time were excluded from the dataset. Statistical analysis was performed using the Kruskal-Wallis non parametric H test: ***p < 0.001. p-values are detailed in Supplementary Table 1.