Fig. 6: DCX reduces neurite numbers by increasing tubulin polyglutamylation levels.

A Images representative of 3 experiments showing CTRL1 and DCXKO4 DIV3 neurons electroporated with DCX-GFP and stained for polyglutamylation. The arrowheads are showing the same cell in both channels (GFP, and polyE). B Violin plots showing the polyglutamylation (polyE) intensity normalized by Tubb3 intensity in CTRL and DCXKO DIV3 neurons with (+ or blue background) or without (− or white background) DCX-GFP electroporation. C Violin plot showing the neurite density in CTRL and DCXKO DIV3 neurons with or without DCX-GFP electroporation. For (B, C), the analysis included n = 97 CTRL0 (light gray), 178 CTRL1 (dark gray), 225 DCXKO1 (cyan), 171 DCXKO4 (dark cyan), 11 CTRL0 + DCX-GFP, 34 CTRL1 + DCX-GFP, 38 DCXKO1 + DCX-GFP, and 45 DCXKO4 + DCX-GFP neurons from 3 experiments. These two plots were cut-off to better show the main population, but the maximum values for polyE intensity, and neurite density are 9.4 (arb. units), 0.156 neurite/µm respectively. Statistical analysis was performed using the Mann-Whitney non parametric U test: ns nonsignificant, **p < 0.01, ***p < 0.001. p-values are detailed in Supplementary Table 3. D Images representative of 2 experiments showing CTRL0 and DCXKO1 DIV3 neurons stained for polyglutamylation (polyE) after 48 h in regular media or 48 h in media supplemented with 100 nM paclitaxel. E Violin plots showing the polyglutamylation (polyE) intensity in CTRL0 and DCXKO1 DIV3 neurons with (+ or green background) or without (− or white background) treatment with paclitaxel. F Violin plots showing the neurite density in CTRL0 and DCXKO1 DIV3 neurons with or without treatment with paclitaxel. For (E, F), images included in the analysis are from n = 84 CTRL0, 172 DCXKO1, 120 CTRL0+paclitaxel and 160 DCXKO1 + paclitaxel neurons from 2 experiments. Statistical analysis was performed using the Mann-Whitney non parametric U test: *p < 0.05, ***p < 0.001. p-values are detailed in Supplementary Table 3.