ZFIN ID: ZDB-FIG-180125-7
Lu et al., 2017 - Overexpression of SOX4 correlates with poor prognosis of acute myeloid leukemia and is leukemogenic in zebrafish. Blood cancer journal   7:e593 Full text @ Blood Cancer J
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: 20-25 somites to Day 5
PHENOTYPE:
Fish:
Observed In:
Stage: Long-pec

Fig. 2

Generation and characterization of the Tg(spi1:SOX4-EGFP) transgenic zebrafish. (a) Schematic diagram showing the LR recombination reaction used to generate the expression constructs, including three entry clones (p5E-spi1, pME-SOX4 and p3E-EGFPpA) and a destination vector (pDestTol2CG2) that contains the cmlc2:EGFP-pA expression cassette. The final construct (pTolCG-spi1:SOX4-EGFP) is shown at the bottom of the figure. (b) Results of semiquantitative PCR showing the expression of SOX4 in TGs and wild-type embryos at 20, 48 and 72 h post fertilization (HPF) and at 5 days. Negative control: non-template; Positive control: pTolCG-spi1:SOX4-EGFP plasmid. (c) Whole-mount immunohistochemical staining (IHC) showed EGFP-positive cells in the heart (yellow arrow) as well as around the yolk sac and caudal hematopoietic tissue (CHT) at 48 HPF. The panel shows GFP+ cells at a higher magnification. (d) Tissue sections showing SOX4-positive cells in the KM of Tg(spi1:SOX4-EGFP) fish but not in those of wild-type fish. The panel shows SOX4-positive cells at a higher magnification. (e) Results from quantitative reverse transcription-PCR (RT-PCR) analysis of hematopoietic marker genes in SOX4 transgenic zebrafish embryos and in wild-type fish embryos. Data are presented as the mean±s.e.m. from three independent experiments. (f, g) Results from whole-mount in situ hybridization (WISH) of mpo and Sudan black (SB) staining showing mpo-positive cells in CHT at 48 HPF and SB-positive cells in CHT at 72 HPF, respectively (magnification: × 40). The panel shows positive cells at a higher magnification (× 100), respectively. Quantification of mpo-positive cells (h) and SB-positive cells (i). Differences among variables were assessed using Student’s t-test.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cebpa AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
csf1ra AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
EGFP twu104Tg (AB) standard conditions Long-pec caudal hematopoietic tissue IFL
Long-pec yolk syncytial layer IFL
gata1a AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
lcp1 AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
mpeg1.1 AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
mpx AB standard conditions Long-pec caudal hematopoietic tissue ISH
Long-pec ventral wall of dorsal aorta ISH
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
Long-pec caudal hematopoietic tissue ISH
Long-pec ventral wall of dorsal aorta ISH
myb AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
runx1 AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
spi1b AB standard conditions 20-25 somites to Day 5 whole organism RTPCR
twu104Tg (AB) standard conditions 20-25 somites to Day 5 whole organism RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab1-sox4 IHC AB standard conditions Long-pec Not Detected head kidney cell
IHC twu104Tg (AB) standard conditions Long-pec head kidney cell
Phenotype Details
Fish Conditions Stage Phenotype
twu104Tg (AB) standard conditions Long-pec granulocyte normal amount, normal
Long-pec hemopoiesis process quality, normal
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 @ Blood Cancer J