Fig. 4

Fig. 4 Loss-of-function of plcg1 affects granulopoiesis independent of plcg1′s role in angiogenesis and in a cell-autonomous manner. ((A)–(D)) WISH analysis of flk1 and mpx expression. In vegfa morphants, flk1 expression is disrupted at 22 hpf ((A) and (B)) indicating defects in angiogenesis. However the mpx expression has no change ((C) and (D)), suggesting granulopoiesis in the ICM is independent of angiogenesis. ((E)–(H)) WISH analysis of gata1 and pu.1 expression. In vegfa morphants, expression of gata1 ((E) and (F)) and pu.1 ((G) and (H)) are not affected. ((I)–(L)) Rescuing experiment with Tol2(flk1: plcg1-2a-mCherry). Fluorescence microscopy images of the trunk of 22 hpf ldd239 mutants injected at one cell-stage with Tol2(flk1:plcg1-2a-mCherry) transgene driving expression of mCherry ((I), red). Injection of Tol2(flk1:plcg1-2a-mCherry) construct can rescue the vascular defect ((J) and (K)) while it cannot rescue decreased mpx expression (L). ((M)–(P)) Rescuing experiment with Tol2(mpx: plcg1-2a-mCherry). Fluorescence microscopy images of the trunk of 22 hpf ldd239 mutants injected at one cell-stage with Tol2(mpx:plcg1-2a-mCherry) transgene driving expression of mCherry ((M), red). Injection of Tol2(mpx:plcg1-2a-mCherry) construct can rescue the decreased mpx expression ((N) and (O)) while it cannot rescue the deficient angiogenesis (P). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)