Minoxidil causes hematopoiesis failure in zebrafish embryos. A Representative images showing staining for mpl:eGFP protein in 4-dpf Tg(mpl:eGFP) larvae treated with egg water with PTU control and minoxidil. Scale bars: 50 μm. B Quantification of mpl:eGFP+ thrombocytes related to (A). Statistical significance was determined using a Mann–Whitney U-test. Data were combined from 4 biological replicates, mean ± SD, ns, not significant; *P < 0.05; ****P < 0.0001. C WISH of mpl expression in control (left panel) and minoxidil-treated (right panel) larvae at 4 dpf. D Quantification of mpl-positive cells in caudal hematopoietic tissue (CHT) (Student’s t-test; mean ± SD;*P < 0.05). E WISH of mpx expression in control (left panel) and minoxidil-treated (right panel) larvae at 4 dpf. F Quantification of mpx-positive cells in CHT (Mann–Whitney U test; mean ± SD; **P < 0.01). G WISH of mfap4 expression in control (left panel) and minoxidil-treated (right panel) larvae at 4 dpf. H Quantification of mfap4-positive cells in caudal hematopoietic tissue (CHT) (Student’s t-test; mean ± SD; ****P < 0.0001). I WISH of rag1 expression in the 4 dpf control (left panel) and minoxidil-treated (right panel) larvae. J Percentage of rag1-positive cells in caudal hematopoietic tissue (CHT) (Fisher’s exact test, ****P < 0.0001). K WISH of βe1-globin expression in the 4 dpf control (left panel) and minoxidil-treated (right panel) larvae. L Percentage of βe1-positive cells in caudal hematopoietic tissue (CHT) (Fisher’s exact test, ****P < 0.0001). M SB staining in control and minoxidil-treated larvae at 4 dpf. N Statistics result of SB-positive neutrophils in CHT region (Student’s t-test; mean ± SD; *P < 0.05). O WISH of c-myb expression in control (left panel) and minoxidil-treated (right panel) embryos at 60 hpf. P Quantification of c-myb-positive cells in CHT (Student’s t-test; mean ± SD; *P < 0.05). Q WISH of c-myb expression in control (left panel) and minoxidil-treated (right panel) embryos at 3 dpf. R Quantification of c-myb-positive cells in CHT (Student’s t-test; mean ± SD; ***P < 0.001)
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