Fetal HSPC development is impaired in smarca5 mutants. (A) Expression of cmyb and runx1 in the AGM region in smarca5^zko1049a and their siblings at 36 hpf by WISH. The quantification of WISH results is shown (bottom). Scale bars, 100 μm. (B) Expression of cmyb in the CHT region in smarca5^zko1049a and their siblings from 2 dpf to 4 dpf by WISH. The quantification of WISH results is shown (right). Scale bars, 100 μm. (C) Expression of erythroid marker gata1, myeloid marker pu.1, and lymphoid marker rag1 in the CHT or thymus region in smarca5^zko1049a and their siblings at 4 dpf by WISH. The quantification of WISH results is shown (right). Scale bars, 100 μm. (D) The imaging of enhanced green fluorescent protein (EGFP) fluorescence in Tg (hsp70:smarca5-EGFP) embryos at 3 dpf. Heat shock was performed at 2.5 dpf. Scale bars, 200 μm. (E) Protein level of Smarca5 in EGFP⁻ and EGFP⁺ embryos in (D) at 3 dpf by western blot. Heat shock was performed at 2.5 dpf. (F) Expression of cmyb in the CHT region and rag1 in the thymus region in smarca5^zko1049a and their siblings, with or without Smarca5 overexpression from 2 dpf to 3 dpf by WISH. The quantification of WISH results is shown (bottom). Scale bars, 100 μm. Data are means ± SD (A-C, F). Asterisk presents statistical significance (*P < .05, ***P < .001; n.s., not significant). P values were calculated by a 2-tailed, unpaired Student t test. HS, heat shock.