Preliminary investigation of morphant vs. mutant phenotype.
WISH within 17 hpf embryos from an in-cross of snai2+/112Δ displayed that the Snail family members snai1a and snai1b are not differentially expressed in mutant embryos as compared to heterozygote and wild-type siblings (A). However, qPCR in pooled embryonic trunks at 26 hpf showed a different trend: all 3 additional members of the Snail family show increased expression (B). This graph presents the average of two independent experiments in which embryonic heads were removed and genotyped, followed by pooling of trunks of the same genotype. Error bars are SD. Snai2 reverse primer is designed within the mutant deletion, so transcript decrease reflects present of mutant transcript. We also observed the effect on emerging HSCs when the SB MO was injected into mutant embryos and their siblings by observing both WISH for runx1 at 26 hpf (C), and the double positive population in Tg(CD41:GFP/kdrl:mCherry) embryos at 48 hpf. Double positive cells were filtered by the surfaces function on Imaris, quantified, and submitted to statistical testing by a non-parametric t-test on Prism (D). Error bars are SEM. By both analyses, HSC specification was affected by SB MO in all genetic backgrounds. Black arrowheads point to the middle of the aortic runx1 expression. **** represents p<0.0001. WT: Wild-type.