Fig. 2: hcASD gene depletion in vivo causes ENCC migration defects.

a Schematic of phox2b staining in NF stage 40 animals to mark enteric neural crest-derived cells (ENCCs, enteric neuron progenitors) in the gut, circled by a blue dashed line. phox2b also labels the hindbrain region and other migrating vagal neural crest cells. b Unilateral mutants were made by injecting the Cas9 protein and an sgRNA targeting an hcASD gene into one cell at the two-cell embryonic stage. Three days later, injected embryos were stained and ENCC area and gut area was measured on each side of the animal and compared to quantify relative gut area (CRISPR side / Control side) of migration. c Individual CRISPR mutagenesis of hcASD genes syngap1, chd8, scn2a, chd2, or dyrk1a reduce the area of ENCC migration in the gut compared to control mutagenesis of pigmentation gene slc45a2. d Area of gut migration quantification by target gene. Control in gray, hcASD genes in blue. A two-way Kruskal-Wallis test was performed followed by Wilcoxon matched-pairs signed rank test to compare the CRISPR side to the control side within each animal and a Holm-Šídák test to adjust for multiple comparisons. Control (N = 24, padj = 0.9248), syngap1 (N = 38, padj = 0.0046), chd8 (N = 25, padj = 0.001), scn2a (N = 33, padj = 0.001), chd2 (N = 31, padj = 0.0011), dyrk1a (N = 38, padj = 0.0011). All samples are independent biological replicates from the same mating pair. In the box plot, whiskers show minimum and maximum values, box represents the 25th and 75th quartiles, and the center line describes the median. Source data are provided as a Source Data file.