|
Fig. EV6 Mouse Sp + /- cortical neurons exhibit a significant number of axonal swellings. (A) Mouse Sp + /+, Sp + /- and Sp-/- cultured cortical neurons immunolabelled with a βIII-tubulin antibody at different days in vitro (DIV). Pink arrows point at axonal swellings. Scale bars: 50 µm. (B, C) Mean number of axonal swellings per 100 nuclei. At least 2500 neurons from two independent experiments were analysed in unblind manner per condition. Violin Plots; horizontal bars indicate the median ± the 1st and 3rd quartiles. Kruskal–Wallis ANOVA test with Dunn’s post hoc test. P values are displayed on graphs. (D) Primary culture of Sp + /- and Sp-/- cortical neurons immunolabelled at DIV9 with βIII-tubulin antibodies, F-actin probes (Phalloidin, pink) and DAPI (grey). Axonal swellings (pink arrows) of Sp + /- exhibit the same characteristic features as those described in Sp-/- cultures. They are (i) always located close to the growth cone (arrowheads), (ii) their diameter is at least 2 to 3 times larger than the diameter of the axon shaft, (iii) they are always strongly labelled by tubulin antibodies and (iv) are always negative for DAPI staining (asterisk). Scale bars: 25 µm. Source data are available online for this figure