Apigenin increases pigmentation in zebrafish. (A) Zebrafish embryos were treated with various concentrations (1–10 μM) of apigenin for different time periods (35–60 h), and zebrafish viability was assessed. (B) Zebrafish embryos were treated with PTU (0.2 mM) for 6–35 h, followed by apigenin (0, 1, 5, or 10 μM) treatment in the embryo medium for 35– 60h. Zebrafish pigmentation was observed using a stereomicroscope. (C) Approximately 30 embryos were collected and dissolved in lysis buffer. After centrifugation, the melanin pigment was dissolved in 200 μL of NaOH working solution (1 mol/L, 10% DMSO) at 80°C for 2 h, and absorbance was measured at 405 nm. Data are expressed as means ± SEM (n = 3). **p < 0.01 vs. non-treated zebrafish, #p < 0.05 and ##p < 0.01 vs. PTU-treated zebrafish. PTU, propylthiouracil; DMSO, Dimethyl Sulfoxide.
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