ZFIN Image: Cortes et al., 2016, Fig. S2

Image

Figure Caption

Fig. S2

Vitamin D treatment expands HSPC number (related to Figure 2).

A. Representative VDRE:mCherry WISH phenotype of embryos treated with 25(OH)D3 and 1,25(OH)D3 compared to DMSO-treated controls (n>10).

B. FACS analysis of Tg(flk1:GFP/VDRE:mCherry) embryos treated with 1,25(OH)D3 from 12-36hpf showed a significant (****p<0.0001) increase in Flk1+ cells responsive to 1,25(OH)D3 (5 embryos/sample x 4 replicates/condition); error bars, mean± SD).

C. FACS analysis of embryos treated with 1,25(OH)D3 (10μM) indicated a 25% increase in EMPs at 30hpf (**p<0.01) compared to controls (n value and error bars as in S2B).

D. FACS analysis of 1,25(OH)D3-treated Tg(flk1:dsred; EF1:mAG-zGEM(1/100) embryos at 36hpf showed a 2-fold increase (***p<0.001) in proliferating endothelial cells, inclusive of the hemogenic population (5 embryos/sample x 4 replicates/condition); error bars, mean ± SD.

E. FACS analysis of vdra MO-injected Tg(flk1:dsred; EF1:mAG-zGEM(1/100) embryos at 36hpf indicated a significant decrease in proliferation (*p<0.05) in the absence of Vitamin D3/VDR signaling (n value and error bars as in S2D).

F. Representative fluorescent microscopy images of Tg(CD41:GFP) embryos treated with 1,25(OH)D3 (1μM) or calcipotriol (1μM) between 12-36hpf depicting a sustained increase in CD41:GFP⁺ expression in the CHT at 72hpf.

G. FACS analysis at 72hpf showed a significant increase in the percentage of CD41:GFP^lo;Gata1:dsRed^- HSPCs after both 1,25(OH)D3 (*p<0.05) and calcipotriol (**p<0.01) treatment (5 embryos/sample x 4 replicates/condition); error bars, mean ± SD.

Scale bar 100μm.

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 @ Cell Rep.