Figure 4.

Kmt2d knockdown affects NPB formation and NC specification in a dose-dependent manner. (A–D) Embryos were injected with 2.5 or 5 ng mmMO or Kmt2d MO into one blastomere at the two-cell stage. 100 pg lacZ mRNA were used for lineage tracing. At neurula stages, the expression of several NC marker genes was analyzed by whole mount in situ hybridization. Injection of 5 ng Kmt2d MO leads to (A) a broader pax3 expression in the NPB region and (B) a severe reduction of foxd3, (C) slug/snai2 and (D) twist expression in the pre-migratory NC. (E–H) Graphs summarizing the results from three independent experiments for each NC marker gene analyzed in (A–D). Kmt2d loss-of-function affects the expression of (E) pax3, (F) foxd3, (G) slug/snai2 and (H) twist in a dose-dependent manner; ±SEM and the number of analyzed embryos are indicated for each condition. A two-tailed unpaired Student’s t-test was applied.