Fig. 2

The number of HSPCs is reduced in the mutant^cas003 CHT.

(A-J′) The time-course analysis of definitive HSPCs from 36hpf to 4dpf by WISH with c-myb (A, A′, C-E′, F, F′, H-J′) or runx1 (B, B′, G, G′) indicating the HSPCs are decreased from 3dpf in mutant^cas003 embryos comparing to siblings. The penetrance of the indicated phenotype is shown in the bottom left of each panel. (A′-J′) Enlarged views of AGM or CHT regions indicated by the dotted rectangles in the left columns. Black arrows represent the AGM (A′, B′, F′, G′) or CHT (C′-E′, H′-J′); red arrows indicate the thymus. (K) Quantitative PCR analysis of c-myb expression level in the tail region from 2dpf to 5dpf showing the attenuated c-myb expression after 3dpf in mutant^cas003 embryos comparing to siblings. Error bars represent SEM. *, p<0.05; ***, p<0.001. (L-R) Immunofluorescence stain of EGFP in the CHT region in mutant^cas003; Tg(c-myb: EGFP) embryos (P-R) and siblings (M-O) from 3dpf to 5dpf. The statistics result of the ratio of EGFP⁺ cell number in the CHT region of mutant^cas003 embryos to that in the siblings is indicated in L (n>6 embryos are counted in each panel). These results indicate that EGFP⁺ HSPCs begin to be decreased obviously in CHT region in mutant^cas003 embryos comparing to siblings from 4dpf. Scale bars represent 50µm. Error bars represent SEM; ns, no significance; *** represents p<0.001.