|
Fig. 11 The effects of ICI 182,780 (ER antagonist) and vegfa morpholino on formononetin-induced angiogenesis in zebrafish embryos in vivo.
Tg (fli1: EGFP)y1 zebrafish embryos (48 hpf) were treated with (A) 0.1% DMSO (negative control), (B) formononetin (50 µM) (positive control), (C) co-treatment of ICI 182,780 (50 µM) and formononetin (50 µM), (D) ICI 182, 780 (50 µM) alone. All treatments were for 24 h. Tg (fli1: EGFP)y1 zebrafish embryos (1 hpf) were injected with (E) Vegfa MO (2 ng) alone and (F) Vegfa MO (2 ng) injection + formononetin treatment (50 µM, at 48 hpf for 24 h) and were observed at 72 hpf. (G,H) Control images of zebrafish embryos at 1–4 cells stage and 24 hpf after Vegfa MO injection. (I) Data were analyzed by using the Image J software package. Quantitative analysis indicated the total length of SIVs for each group. (J) Evaluation of gene expressions in formononetin-treated zebrafish embryos by using real-time PCR. Data are plotted as means ± SD from three individual experiments. *p < 0.05, **p < 0.01 vs. control group.