Fig. 3

Enpp2 overexpression induces alteration in cellular migration, the expression pattern of KV precursors, dorsal organizer and midline genes (gsc and shha) during early embryogenesis.

(A) Representative images of WISH with sox17 riboprobe to mark the integrity of the DFC cluster at 75% epiboly. (B) Quantification of KV defect and/alteration of DFC cluster, as highlighted by sox17 WISH. (C) The gsc expression is observed in the dorsal organizer of enpp2 overexpressing embryos. However, the expression is also detected in the axial mesoderm precursors suggesting that convergence and cell migration during midline formation is impaired. (D) Representative images of WISH of shha at 90% epiboly showing a truncated axis and laterally expanded expression of shha at the blastomderm margin. (E) Quantification of the broadened shha expression domain at the blastoderm margin in control and enpp2 overexpressing embryos at 90% epiboly. (F,G) Representative images of time lapse series showing cell migration during midline formation (10 hpf- early somitogenesis) in wild type and enpp2-injected embryos. Image stacks were taken every 15 minutes. Time course for each image is indicated. The enpp2 overexpressed embryo failed to undergo a proper CE (G) which results in bend and broadened midline compared to wild type (F). (B,E) The sample size (n) is stated as numerical value above each bar. Data are mean ± SEM. Statistical analysis was established by t-test; *P < 0.05; **P < 0.01. n indicated above bars.