Fig. 4

a–d Representative confocal images of WT (a–c) and AS (d–f) HNs (DIV4), cultured under control conditions (a, d), or treated with nocodazole 40 nM (b, e) or with blebbistatin 25 μM (c, f). Neurons were cultured on GRs, treated with drugs from early DIV2, and immunostained for SMI312 (axonal marker, green) and MAP2 (dendrite marker, red) at DIV4. The underlying GR pattern is reported as insets; inset side = 60 μm. g–i Axonal morphological parameters of WT (white columns) and AS (grey columns) HNs on GRs in the presence of Noco (squared columns) and Bleb (striped columns): axon alignment (g), straightness (h), and mean length (i) were calculated per each cell. g ***P < 0.001 WT vs. AS, *P < 0.05 WT vs. AS+Noco, **P < 0.01 AS vs. AS+Noco and WT+Bleb, Bonferroni’s test; within WT HNs samples: °°P < 0.01 WT vs. WT+Noco and WT+Noco vs. WT+Bleb, Bonferroni’s test; within AS HNs samples: °P < 0.05 AS vs. AS+Noco, Bonferroni test. h *P < 0.05 WT vs. AS and vs. AS+Bleb, Bonferroni’s selected test; within WT HNs samples: °^/°°°P < 0.05–0.001 WT vs. WT+Noco and WT+Bleb, Bonferroni’s test. i P > 0.05, Bonferroni’s test. N ≥ 3, at least 15 HNs for samples