Figure 3.

MEQ inhibits angiogenesis in vivo. (A, B) The embryos of transgenic zebrafish (Tg [Fli1a: EGFP]) at 1-2 cell stage were injected with MEQ and vascular development was monitored at 72 h post-fertilization (A) and the percentage of fish with deficiency in intersegmental vessels (Hypo-ISV) were analyzed (B). ***P < 0.001 using the χ²-test. (C) The qRT-PCR results show the expression of Sars (i.e., fish SerRS) and Vegfa in the zebrafish treated with MEQ or dimethyl sulfoxide (DMSO). Data are plotted as the mean ± SEM (n = 3; *P < 0.05; ***P < 0.001 using Student’s t-test). (D) Western blotting shows SerRS protein levels induced by MEQ in 4T1 cells. (E) Representative images of Matrigel plugs formed by 4T1 cells treated with MEQ or DMSO control. Scale bar represents 1 cm. (F–H) Immunofluorescence of CD31(F), immunochemical staining of SerRS (G) and VEGFA (H) in the Matrigel plugs and the quantification of positively-stained cells are plotted as the mean ± SEM (n = 5; *P < 0.05; ***P < 0.001 using Student’s t-test. Scale bars represent 100 μm.