7,17-diHDPAn-3 promotes HSC formation. (A) WISH for cmyb, labeling definitive HSCs at 36 hpf, in controls and embryos treated with 7,17-diHDPAn-3 from 11 to 36 hpf (n = 9 embryos per group). Images were taken at original magnification ×10. (B) Quantification of WISH signal intensity using Fiji software showed a significant increase in cmyb expression in the treated group compared to controls (n = 5-7 embryos per group; unpaired Student t test). (C) RT-qPCR analysis showed relative expression of cmyb and runx1 in treated embryos vs controls from 11 to 36 hpf (n = 4 biological replicates per group; technical triplicates; 30 embryos per sample). (D) RT-qPCR of dissected heads revealed cmyb and runx1 expression after exposure from 11 to 36 hpf (n = 3 biological replicates; technical triplicates; 12 heads per sample). (E) RT-qPCR of dissected tails revealed cmyb and runx1 expression after exposure from 11 to 36 hpf (n = 3 biological replicates; technical triplicates; 12 tails per sample). Bar plots represent mean ± standard deviation (SD). Statistical comparisons between control and treated groups for each gene were performed using unpaired Student t tests. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. ctrl, control; ns, no significant difference.
Figure 1.

7,17-diHDPAn-3 promotes HSC formation. (A) WISH for cmyb, labeling definitive HSCs at 36 hpf, in controls and embryos treated with 7,17-diHDPAn-3 from 11 to 36 hpf (n = 9 embryos per group). Images were taken at original magnification ×10. (B) Quantification of WISH signal intensity using Fiji software showed a significant increase in cmyb expression in the treated group compared to controls (n = 5-7 embryos per group; unpaired Student t test). (C) RT-qPCR analysis showed relative expression of cmyb and runx1 in treated embryos vs controls from 11 to 36 hpf (n = 4 biological replicates per group; technical triplicates; 30 embryos per sample). (D) RT-qPCR of dissected heads revealed cmyb and runx1 expression after exposure from 11 to 36 hpf (n = 3 biological replicates; technical triplicates; 12 heads per sample). (E) RT-qPCR of dissected tails revealed cmyb and runx1 expression after exposure from 11 to 36 hpf (n = 3 biological replicates; technical triplicates; 12 tails per sample). Bar plots represent mean ± standard deviation (SD). Statistical comparisons between control and treated groups for each gene were performed using unpaired Student t tests. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. ctrl, control; ns, no significant difference.

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