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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: Exp Hematol. 2016 Oct 15;46:83–95.e6. doi: 10.1016/j.exphem.2016.10.002

Fig 3. PDGF-B stimulation increases developmental HSPC numbers.

Fig 3

(A) Overexpression of pdgfb by mRNA injection enhanced runx1;cmyb expression in the VDA by WISH at 36hpf.

(B) Qualitative phenotypic distribution of embryos from panel 3A (n≥20/condition × 5 replicate clutches).

(C) FACS analysis confirmed that pdgfb overexpression increased Flk1:dsRed+cMyb:GFP+ HSPCs at 48hpf (1.54 fold increase, ***p≤0.0001, two-tailed t test, n≥5 replicates/condition).

(D) WISH analysis showed enhanced cmyb expression at 48hpf following ectopic activation of PDGFRβ signaling using hsp70:ca-pdgfrb induced at 27hpf (37°C for 1 hour)

(E) Qualitative phenotypic distribution of embryos from panel 3D (n≥20/condition × 5 replicate clutches).

(F) FACS analysis confirmed a significant increase in Flk1+cMyb+ HSPCs (48hpf) following PDGFRβ activation mediated by induced expression of ca-pdgfrb (1.47 fold increase, *p<0.05, two-tailed t test, n≥5 replicates/condition).

(G) Overexpression of pdgfb ligand rescued the reduction in runx1;cmyb expression found in hif1a morphants at 36hpf.

(H) Qualitative phenotypic distribution of embryos from panel 3G (n≥20/condition × 3 replicate clutches).