Definitive hematopoiesis is impaired in the smu1347 mutants. A, B, E, F WISH showed that expression of βe1 (A), lyz (E), and mfap4 (F) was markedly reduced in the CHT region in the smu1347 mutants at the corresponding stage. rag1 (B) expression was nearly absent in the thymus of smu1347 mutants at 5 dpf. Red circles indicated signals in the thymus region. The red and blue arrows denoted microcephaly and microphthalmia, respectively, in smu1347 mutants. C, D, G, H Quantification of βe1+ signal area, lyz+, mfap4+ cell numbers in the CHT region and rag1+ signal area in the thymus revealed significant reductions in smu1347 mutants (mean ± SD; n > 10; Student’s t test and nonparametric Mann–Whitney test, ****p < 0.0001). I WISH of cmyb showed that cmyb+ HSPCs were comparable at 2 dpf but significantly diminished in the CHT region of the smu1347 mutants by 3–4 dpf. Red arrows indicated relevant signals in the region. J Quantification of cmyb+ cell numbers in the CHT region at corresponding stages of siblings and smu1347 mutants (mean ± SD; n ≥ 10; Student’s t test and nonparametric Mann–Whitney test, ns p > 0.05, ***p < 0.001, ****p < 0.0001). Data from three independent experiments were pooled for analysis. The ratio in the upper right corner of WISH data indicated the proportion of embryos with representative expression pattern for each genotype. Scale bars: 200 μm.
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