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Fig. 3 Effects of IXA4 and TUDCA on survival, hatching rate, and morphology in zebrafish embryos and larvae exposed to Tl.(A) Survival rates of zebrafish embryos exposed to Tl (200 μg/L) with or without IXA4 (20 μM) or TUDCA (20 μM) treatments to 144 hpf. Data are presented as mean ± SD (n = 10–15 embryos per group). (B) Hatching rates of zebrafish embryos at 60 hpf under different treatment conditions, including control, IXA4 (20 μM), TUDCA (20 μM), Tl (200 μg/L), Tl + IXA4, and Tl + TUDCA. Data are expressed as mean ± SD (n = 10–15 embryos per group). (C) Representative images of zebrafish larvae at 144 hpf under control conditions or exposed to Tl (200 μg/L) with or without IXA4 (20 μM) or TUDCA (20 μM) treatments, showing morphological changes across treatment groups. (D) Representative images of zebrafish larvae at 144 hpf exposed to Tl (200 μg/L) with or without IXA4 (20 μM) or TUDCA (20 μM). Morphological abnormalities included swim bladder deflation (blue arrows), yolk retention (blue dashed-line arrows), and pericardial edema (black arrows). (E) Morphology scores of zebrafish larvae at 144 hpf exposed to Tl with or without IXA4 or TUDCA. Data are presented as individual data points for each group, with horizontal lines indicating the mean values. (F) Swim bladder area (mm²) and (G) abdominal area (mm²) measurements of zebrafish larvae at 144 hpf. Data are presented as individual data points for each group. Statistical significance was determined using t-tests. * *p < 0.01, * **p < 0.001, compared to the indicated group. n = 10–15 embryos per group. Scale bar, 500 µm.