Definitive myelopoiesis is strongly expansive in pten-deficient embryos. (A-B) Whole-mount in situ hybridization analyses of definitive myelopoiesis with riboprobes for lyz in the indicated embryos at 90 hpf. The lower panels are high-magnification views of the indicated region in the upper panel. The red arrowheads indicate myeloid cells in the CHT and kidney/thymus region. (C-D) The pten-/-lyz:EGFP embryos show increased EGFP-positive myeloid cells. The red arrowheads indicate EGFP-positive cells in the CHT and kidney region. The embryos are shown in lateral views with the anterior to the left. e: eye. (E-F) Statistical result for a, b and c, d respectively. (G-M) Whole-mount in situ hybridization analyses for other myeloid markers (l-plastin, mpo, nephrosin) in the CHT. The red arrowheads indicate the myeloid cells marked by each probe. The data shown in (E-F) and (M) are the means ± SEM of at least 30 embryos; ***p < 0.001 versus the control. Scale bar: 100 μm.