|
Fig. 5 Gata5/6 and Hedgehog signaling are required for endocardial myelopoiesis (A–D) Co-injection of gata5 and gata6 MOs results in a complete or nearly complete absence of spi1b- (B) or lcp1-positive cells (D) compared with control uninjected embryos (A and C). In situ hybridization analysis of spi1b and lcp1 expression at the 24-hpf stage is shown. Scale bar, 100 μm. (E–J) Ventral view of the cardiac region of control and smo−/− mutant embryos processed for myeloid spi1b FISH (green) and myocardial (myoc.) MF20 immunostaining (red) at 24 hpf. Maximum-intensity projection is shown. Scale bar, 50 μm. (K) Quantification of cell number inside and outside the heart tube in control (n = 22) and smo−/− mutant (n = 24) embryos. Student’s t test, error bars show ±SD. NS, non-significant.