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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 2. PDGFRβ signaling acts downstream of Hif1α to control AGM HSPC production.

Fig 2

(A) Morpholino knockdown of pdgfrb attenuated the increase in runx1;cmyb expression in embryos treated with the Hif1α agonist DMOG (75μM).

(B) Qualitative phenotypic distribution of embryos from panel 2A (n≥20/condition × 3 replicate clutches).

(C) Knockdown of vhl increased runx1;cmyb WISH expression in the AGM at 36hpf, while co-injection with the pdgfrb MO blocked this effect.

(D) Qualitative phenotypic distribution of embryos from panel 2D (n≥20/condition × 2 replicate clutches).

(E) FACS analysis for CD41+(Gata1−) HSPCs following injection of MOs to vhl and pdgfrb alone and combined confirmed a reduced impact for Hif1α stabilization with loss of PDGF signaling (*p<0.05, one-tailed t-test, n≥7).

(F) Ectopic activation of a dominant negative pdgfrb transgene, Tg(hsp70:dn-pdgfrb), at 27hpf (37°C for 1 hour) diminished the impact of CoCl2 stimulation on cmyb expression at 48hpf.

(G) Qualitative phenotypic distribution of embryos from panel S2G (n≥20/condition × 4 replicate clutches).

(H) CD41+ HSPC cell counts at 48hpf confirmed the effect of loss of PDGFRB signaling, mediated by induction of a dominant negative receptor, on Hif1α activation by CoCl2 (*p<0.05, **p<0.01, one-tailed t-test, n≥8).