ZFIN ID: ZDB-FIG-181004-20
Bickers et al., 2018 - Zebrafish snai2 mutants fail to phenocopy morphant phenotypes. PLoS One   13:e0202747 Full text @ PLoS One
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 9

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.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
runx1 WT standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
WT + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
snai2sd57/+ standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
snai2sd57/+ + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
snai2sd57/sd57 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
snai2sd57/sd57 + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell ISH
snai1a snai2sd57/sd57 standard conditions Prim-5 trunk RTPCR
snai1b snai2sd57/sd57 standard conditions Prim-5 trunk RTPCR
snai2 snai2sd57/sd57 standard conditions Prim-5 trunk RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell runx1 expression decreased amount, abnormal
hzm3Tg; s896Tg; sd32Tg + MO1-snai2 standard conditions Long-pec ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
snai2sd57/+ + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell runx1 expression decreased amount, abnormal
snai2sd57/+; hzm3Tg; s896Tg; sd32Tg + MO1-snai2 standard conditions Long-pec ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
snai2sd57/sd57 standard conditions Prim-5 trunk snai1a expression increased amount, abnormal
Prim-5 trunk snai1b expression increased amount, abnormal
Prim-5 trunk snai2 expression decreased amount, abnormal
snai2sd57/sd57 + MO1-snai2 standard conditions Prim-5 ventral wall of dorsal aorta hematopoietic stem cell runx1 expression decreased amount, abnormal
snai2sd57/sd57; hzm3Tg; s896Tg; sd32Tg + MO1-snai2 standard conditions Long-pec ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS One