ZFIN ID: ZDB-FIG-161111-5
Cortes et al., 2016 - Developmental Vitamin D Availability Impacts Hematopoietic Stem Cell Production. Cell Reports   17:458-468 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Conditions:
Anatomical Term:
Stage: Prim-25
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Prim-25 to Long-pec

Fig. 2

1,25(OH)D3 Regulates HSPC Numbers via Vitamin D Receptor Signaling

(A) Representative runx1/cmyb WISH images and qualitative phenotype distribution following 25(OH)D3 exposure from 12–36 hpf (n ≥ 40 embryos/condition).

(B) FACS analysis of 25(OH)D3-treated (10 μM) embryos showed a significant increase in HSPCs (∗∗p < 0.01) (5 embryos/sample × 4 replicates/condition). Error bars, mean ± SD.

(C) qPCR analysis indicated a significant induction in the VDR transcriptional target cyp24a1 at 36 hpf after exposure to 25(OH)D3 (p < 0.05) or 1,25(OH)D3 (∗∗∗p < 0.001) (12–36 hpf) (30 pooled embryos/condition × 3 replicates).

(D) Phenotype distribution of runx1/cmyb expression in embryos exposed to increasing concentrations of 1,25(OH)D3 from 12–36 hpf (n ≥ 30 embryos/condition).

(E) FACS analysis of 1,25(OH)D3-treated (10 μM) embryos showed a 20% increase in HSPCs at 48 hpf (∗∗p < 0.01) compared with controls (n value and error bars as in B).

(F) runx1/cmyb WISH and phenotype distribution after exposure to the non-calcemic vitamin D3 analog calcipotriol (n ≥ 30 embryos/condition).

(G) FACS analysis of calcipotriol-treated embryos showed a significantly more HSPCs (p < 0.05) at 36 hpf (n value and error bars as in B).

Scale bars, 100 μm. See also Figure S2.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
cyp24a1 WT control Prim-25 whole organism RTPCR
WT chemical treatment by environment: calcitriol Prim-25 whole organism RTPCR
WT chemical treatment by environment: calcidiol Prim-25 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: calcitriol Prim-25 hematopoietic stem cell proliferation increased occurrence, abnormal
Prim-25 ventral wall of dorsal aorta has extra parts of type hematopoietic multipotent progenitor cell, abnormal
Prim-25 whole organism cyp24a1 expression increased amount, abnormal
WT chemical treatment by environment: calcidiol Prim-25 hematopoietic stem cell proliferation increased occurrence, abnormal
Prim-25 ventral wall of dorsal aorta has extra parts of type hematopoietic multipotent progenitor cell, abnormal
Prim-25 whole organism cyp24a1 expression increased amount, abnormal
pd27Tg; sd2Tg; zf169Tg; zf71Tg chemical treatment by environment: calcitriol Long-pec hematopoietic stem cell proliferation increased occurrence, abnormal
Long-pec ventral wall of dorsal aorta has extra parts of type hematopoietic multipotent progenitor cell, abnormal
pd27Tg; sd2Tg; zf169Tg; zf71Tg chemical treatment by environment: calcidiol Prim-25 hematopoietic stem cell proliferation increased occurrence, abnormal
Prim-25 ventral wall of dorsal aorta has extra parts of type hematopoietic multipotent progenitor cell, abnormal
pd27Tg; sd2Tg; zf169Tg; zf71Tg chemical treatment by environment: calcipotriol Prim-25 hematopoietic stem cell proliferation increased occurrence, abnormal
Prim-25 ventral wall of dorsal aorta has extra parts of type hematopoietic multipotent progenitor cell, abnormal
Acknowledgments:
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