Fig. 7

Snai2^112Δ and snai2^sa24539 mutants have no embryonic defects in HSC or sclerotome formation.

WISH analysis was performed on embryos derived from heterozygote in-crosses including probing for the hematopoietic markers runx1 (A) and cmyb (B) and the sclerotome marker foxc1b (C). Wild-type, heterozygotes, and mutants are all included for the Snai2^112Δ allele since genotyping without sequencing was possible. For the snai2^sa24539 allele, genotyping by PCR was sufficient to determine which embryos lacked the wild-type SNP, but not to distinguish wild-type from heterozygote. Thus, images are included for mutants versus “siblings”. For all markers, no obvious defect is detected. Black arrowheads point to the middle of the aortic runx1 or cmyb expression. Numbers in the lower right-hand corner of each image depict the number of embryos with the phenotype pictured out of the total number of embryos assayed in each condition. HSC specification was also analyzed in Tg(CD41:GFP/kdrl:mCherry) fish on the snai2^112Δ background by confocal microscopy at 48 hpf, and Imaris imaging software was used to remove GFP signal outside of the vasculature (D). Pink coloration is indicative of double positive cells as filtered by the surfaces feature of Imaris. White arrowheads indicate separate putative HSPCs, and quantification for each fish was graphed and statistically analyzed by a non-parametric t-test on Prism (E). Error bars are SEM.