CH₂Cl₂ extract reduces total visceral adipose tissue (VAT) in high‐fat diet‐fed zebrafish. (a) Experimental design of this study. (b) Preparation of MO extracts. Extracts were prepared using serial extraction with each solvent. (c) NR fluorescent intensities in the VAT were calculated. The Y‐axis indicates the ratio of NR staining before and after 48‐h treatment with each extract. Data are shown as the means ± standard deviation. n = 5, *p < .05 versus control, as calculated by using one‐way ANOVA. Reproducibility test results are displayed in Figure S2. Each extract was administered at 10 μg/ml. Phenylephrine (20 μM) was used as a positive control. (d) Representative images of control‐ and CH₂Cl₂ extract‐treated zebrafish

Fraction 2 of CH₂Cl₂ extract suppresses adipogenesis in zebrafish VAT. (a) Flow chart showing elution fractions obtained using silica gel chromatography. (b and c) Zebrafish obesogenic test. Fraction 2 (10 μg/ml) reduced the amount of VAT compared to control treatment (0.1% DMSO). Data are shown as the means ± standard deviation. n = 5, *p < .05 versus control, as calculated by using one‐way ANOVA. Reproducibility test results are provided in Figure S3. (d and e) qPCR analysis of early adipocyte differentiation markers CCAAT/enhancer‐binding protein beta (cebpb) and CCAAT/enhancer‐binding protein delta (cebpd; d) and late differentiation markers peroxisome proliferator‐activated receptor gamma (pparg) and CCAAT/enhancer‐binding protein alpha (cebpa; e). Data are shown as the means ± standard deviation. n = 5 or 6, **p < .01 versus control, as calculated by using the Student's t‐test