Screening of in vivo estrogenic activity with transgenic medaka. (A) Bakuchiol induced GFP expression in liver, the white bar indicates 100 µm. (B) LC₅₀ and EC₅₀ of fish after exposure to bakuchiol for 24 h. Correlation between different concentrations of (C) bakuchiol and (D) E2, and the induced GFP signal intensity. (E) Analysis of mRNA expression of ERα and ERβ by using real-time PCR. Fish (n = 10) were exposed to 0.5 µg/ml bakuchiol for 24 h. 18S rRNA was used as the internal reference. BAK, bakuchiol.

Bakuchiol inhibited in vivo cell mass formation in zebrafish. (A) ~300 MCF-7 cells were injected into the yolk sac of 48 hpf zebrafish embryos. 60 embryos were treated with ethanol (solvent control) or 0.5 µg/ml of bakuchiol for 72 h. Triplicates for each condition were performed. Representative bright field and fluorescent images of embryos in both control and treatment groups showing cell mass in the yolk sac area were shown. The black and white bars indicate 500 µm. (B) Statistical analysis of integrated fluroscent intensity of cell mass in control and treatment group (*p < 0.05, one sample t-test). Ctrl, control; BAK, bakuchiol.