Fig. 7: Conditional deletion of Trio in GABAergic INs disrupts their ability to respond to the guidance and motogenic cues Netrin-1 and EphA4, leading to impaired IN migration.

Graphs showing the normalized migratory index of dissociated Trio^WT and Trio^cKO MGE-INs in Boyden chamber assays following treatment with vehicle or Slit-2 (A), or with vehicle or Netrin-1 (B). (n = 5 experiments for Trio^WT and 6 experiments for Trio^cKO MGE-INs treated with vehicle, Netrin-1 or Slit-2). Examples of fluorescence images of Trio^WT and Trio^cKO MGE explants cultures 48 h after treatment with either vehicle (C, E) or Netrin-1 (D, F). Dotted lines illustrate the migration radius on each explant. G Violin-box graph showing the normalized migration radius index of MGE-INs migrating out of Trio^WT and Trio^cKO explants 48 h after treatment with Netrin-1. (n = 11 MGE explants from 3 Trio^WT brains and 13 MGE explants from 3 Trio^cKO brains treated with vehicle and 26 MGE explants from 3 Trio^WT brains and 16 MGE explants from 3 Trio^cKO brains treated with Netrin-1). H Graph showing the normalized migratory index of dissociated Trio^WT and Trio^cKO MGE-INs in Boyden chamber assays following treatment with human protein FC or FC-EphA4. (n = 4 experiments for Trio^WT and 3 experiments for Trio^cKO MGE-INs treated with human FC or FC-EphA4 protein). Examples of fluorescence images of Trio^WT and Trio^cKO MGE explants cultures 48 h after treatment with either human protein FC (I, K) or FC-EphA4 (J, L). M Violin-box graphs showing the normalized migration radius index of MGE-INs migrating out of Trio^WT and Trio^cKO explants 48 h after treatment with human protein FC or FC-EphA4 (n = 17 MGE explants from 4 Trio^WT brains and 30 MGE explants from 6 Trio^cKO brains treated with human FC protein and 18 MGE explants from 4 Trio^WT brains and 28 MGE explants from 6 Trio^cKO brains treated with human FC-EphA4). Violin-box graphs showing Trio^WT and Trio^cKO MGE-INs migration dynamics following treatment with human protein FC or FC-EphA4, including nucleokinesis frequency (N), pause duration (O), total migration velocity (P), net displacement (Q), total traveled distance (R) and nucleokinesis amplitude (S), duration (T) and velocity (U). (n = 32 MGE-INs from 3 Trio^WT brains and 29 MGE-INs from 3 Trio^cKO brains treated with human FC protein and 34 MGE-INs from 3 Trio^WT brains and 28 MGE-INs from 3 Trio^cKO brains treated with human FC-EphA4). *P < 0.05, **P < 0.01, ****P < 0.0001 by two-way ANOVA followed by Bonferroni’s multiple comparisons test. For better clarity, only comparisons between treatments for each genotype were made. Scale bars: 500 µm.