7,17-diHDPAn-3 modulates inflammatory signaling and promotes HSC formation. (A) RT-qPCR analysis of inflammatory cytokine genes (il1b, il6, il8, tnfa, and il10). (B) WISH analysis of cmyb expression at 36 hpf in zebrafish embryos treated with 75 μM dexamethasone (11-36 hpf), compared to controls. Representative images shown at 10× magnification (n = 9 embryos per group). (C) Quantification of cmyb WISH signal in the AGM using Fiji software confirms a significant increase in dexamethasone-treated embryos (n = 4 biological replicates). (D-E) RT-qPCR analysis of cmyb and runx1 expression across a dexamethasone dose-response series: 37.5 μM (D) and 75 μM (E). (F) RT-qPCR quantification of cmyb and runx1 expression after 7,17-diHDPAn-3 treatment. (G) RT-qPCR analysis of embryos treated with subthreshold concentrations of 18.75 μM dexamethasone or 0.375 μM 7,17-diHDPAn-3 individually shows no significant increase in cmyb or runx1 expression. (H) Combined treatment with 18.75 μM dexamethasone and 0.375 μM 7,17-diHDPAn-3 increases cmyb and runx1 expression, indicating a potential cooperative effect. RT-qPCR was performed using 3 biological replicates with technical triplicates; 30 embryos per biological replicate. Data are presented as mean ± SD. Statistical analysis was performed using an unpaired Student t test. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. ctrl, control; ns, no significant difference.