Fig. 3

Muscone abrogates tumor angiogenesis in cellular and animal models. A, B Tube formation in vitro of HUVECs treated with muscone (A), or TCM from muscone-treated MDA-MB-231 cells (231TCM) or BT-549 cells (549TCM) (B). Tube lengths were measured with Fiji imaging software; Scale bar, 100 µm. C Equal numbers of Dil-labelled and incorporated MDA-MB-231 cells were administered into the zebrafish embryos perivitelline space at 48 h after fertilization. Arrows mark the administration locations. Magnification: 28.5 × . D Vasculature and neo-angiogenesis around the primary tumor and metastasis locations in zebrafish treated with muscone for 48 h. Boxes indicate newly formed tumor vessels. Magnification: 100 × . E Photographs of the Matrigel plugs from mice treated with VEGF and muscone for 12 d. F Representative images of immunofluorescent CD31⁺ microvessels (red) within Matrigel plugs; Scale bar, 100 µm. G Microvessel densities quantified as the density of CD31⁺ cells. H The effect of TCM from muscone-treated MDA-MB-231 cells (231TCM) on tumor angiogenesis-related gene expression in HUVECs. All data are shown as mean ± SEM, n = 3, *P < 0.05, **P < 0.01, and ***P < 0.001