FIGURE

Fig. S5

ID

ZDB-FIG-161111-9

Publication

Cortes et al., 2016 - Developmental Vitamin D Availability Impacts Hematopoietic Stem Cell Production

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Fig. S5

Expression of Cxcl8 receptors cxcr1/cxcr2 in zebrafish HSPCs (related to Figure 5).

A. Expression analysis of FACS sorted populations showed enhanced cxcl8 expression following exposure to 1,25(OH)D3, with the biggest effect in Flk1⁺/cMyb⁺ HSPCs (cells sorted from ≥1000 embryos/condition x 2 replicates); error bars, mean ± SD.

B. qPCR analysis of ZFcxcr1 and Zfcxcr2 expression shows enrichment of ZFcxcr1 in FACS sorted HSPCs (Flk1⁺/cMyb⁺) and the Flk-/cMyb- population (n=2 replicates, cells sorted from ≥1000 embryos/condition); N.D. (not detected).

C. WISH analysis and phenotype distributions of embryos treated with 1,25(OH)D3 with and without the CXCL8 inhibitor SB25002 (1μM) showed co-treatment attenuated Vitamin D-mediated increases in runx1/cmyb expression (n≥50 embryos/condition).

D. FACS quantification of Flk1⁺/cMyb⁺ HSPCs after exposure to 1,25(OH)D3 with and without SB25002 (Con vs 1,25(OH)D3, *p<0.01; 1,25(OH)D3 vs SB25002, *p<0.01; Con vs 1,25(OH)D3 + SB25002, N.S.; (5 embryos/sample x 4 replicates/condition; error bars, mean ± SD ).

Scale bar 100μm.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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