Fig. 4

Emb is required for the migration of the ENCCs in gut explant. A, Schematic illustrating the ENCC migration assay in mouse gut explants. B Using the transgenic mice with ENCCs labeled by genetically encoded tdTomato, and the ENCCs were marked by Sox10 (red) in E12.5d mice. C, Relative density of ENCCs behind the leading ENCCs, NS, non-significance, one-way ANOVA with Bonferroni post-hoc, N = 4. D, Representative images of the in vitro ENCC migration assay, ENCCs are labeled with P75NTR (red) and Sox10 (green). E, Quantification methods of the in vitro migration assay, the membranes were divided into five regions based on the distance from the margin of the explant and the numbers of Sox10 positive cells in each region were counted and quantified. F and G, Mean migration distance of Sox10+ cells and migration distance of the furthest migrating Sox10+ cell in different groups; **, p < 0.01; ***, p < 0.001, one-way ANOVA with Bonferroni post-hoc, N = 3. H and I, Polar histograms representing the trajectories of the cells which were 150 μm from the most caudal cell at 40-min intervals in WT (H) and Emb-/- explants of E12.5d hindgut (I). The trajectories are determined by drawing a straight line from the position of cell to its position 40 min previously, with the 0 degree as the rostrocaudal axis of the gut (27 cells were examined). J, Quantification of the average net speed by measuring the distance between the location of the wave front at the beginning of the time lapse sequence and its location at the end of the sequence. ***, p < 0.001, two-tailed Student’s T test, N = 3