7,17-diHDPAn-3 increases erythroid progenitor production. (A) WISH for gata1, labeling erythroid progenitors at 18, 24, and 36 hpf, in embryos treated with 7,17-diHDPAn-3 from 11 hpf, vs controls (n = 9 embryos per group per time point). Images were taken at ×5 magnification (18-hpf embryos) and original magnification ×10 (24-hpf and 36-hpf embryos). (B) Quantification of WISH signal intensity using Fiji software showed a significant increase in gata1 expression in treated embryos at all time points compared to controls (n = 5-7 embryos per group; unpaired Student t test). (C-D) RT-qPCR analysis showed relative expression of gata1 in embryos treated from 11 to 18 hpf (C) and from 11 to 36 hpf (D) vs controls (n = 4 biological replicates per group; technical triplicates; 30 embryos per sample). Bar plots represent mean ± SD. Statistical comparisons between the control and treated groups for each gene were performed using unpaired Student t tests. ∗P < .05; ∗∗P < .01. ctrl, control.