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Fig. 3

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ZDB-IMAGE-151009-26
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Figures for Jia et al., 2015
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Fig. 3

Transient expression of Bcl2 rescues the defective hematopoiesis in kri1lcas002 mutant embryos. (A) Statistic analysis of the proportion of proliferative HSPCs (cmyb and PH3 double positive) in total HSPCs in kri1lcas002 mutant embryos and wild-type siblings during hematopoietic development (40 hpf-2 dpf). N e 10 for each group. 40 hpf AGM, p = 0.0873; 40 hpf CHT, p < 0.0001; 2 dpf CHT, p<0.0001. Error bars represent SEM. **P ≤ 0.01; ***P ≤ 0.001 (Student′s t-test). (B) Proportion of kri1lcas002 mutant embryos with classified hematopoiesis phenotype (indicated by cmyb staining) with or without microinjection of p53 MO, treatment with caspase inhibitor (Z-VAD-FMK) or Bcl2 expression, respectively. (C-E) The CHT hematopoiesis is recovered by Bcl2 overexpression in kri1lcas002 mutant embryos. Embryos from two heterozygous parents were injected with bcl2 mRNA at one-cell stage and harvested for WISH analysis. After WISH and photographing, all embryos were genotyped by genomic DNA sequencing; percentage of rescue was then calculated. 26 out of 33 mutant embryos injected at one-cell stage with bcl2 mRNA are fully rescued, while the rest of embryos are partially rescued. (C′-E′) The enlarged CHT regions. (F) Percentage of the proliferative HSPCs in the CHT of kri1lcas002 mutant embryos and wild-type siblings with or without Bcl2 overexpression at 3 dpf. N = 12 for each group. Without Bcl2 overexpression, p < 0.0001; With Bcl2 overexpression, p = 0.3136. Error bars represent SEM. **P ≤ 0.01; ***P ≤ 0.001 (Student′s t-test). (G-R) Representative confocal images of double staining of cmyb RNA (red) and pH3 protein (green) in the CHT at 3 dpf. G, K and O are bright-field images overlaid with fluorescent staining. Scale bars, 50 µm.

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