Fig. 7

NONO knockdown inhibits the migration and invasion potential of breast cancer cells invitro and in vivo. a, b The migration ability of siRNA-scr and siRNA-NONO transfected breast cancer cells was investigated using a wound healing assay. ImageJ 64 was used to measure wound area. Magnification, 10X. c The Transwell Migration and Invasion Assay determined the migratory and invasive capacities of MDA-MB-231 and MCF-7 breast cancer cells transfected with siRNA-scr and siRNA-NONO. Scale bar, 100 µm and magnification, 20X. a-c Data are representations of three independent experiments. *p < 0.05, **p < 0.01, p*** < 0.001. d Western blot analysis of the expression of E-cadherin, vimentin, MMP-2 and MMP-9 proteins in the siRNA-scr and siRNA-NONO-transfected breast cancer cells. β-actin served as a protein loading control. The epithelial marker E-cadherin was upregulated, and the mesenchymal marker vimentin was downregulated in NONO knockdown cells. Endogenous MMP-2 and MMP-9 expression levels were reduced by NONO knockdown. Representative blots from at least two independent experiments are shown. β-actin was used as an internal control. e The migration of siRNA-scr and siRNA-NONO-transfected MDA-MB-231 cells was studied by staining the cells with Green CMFDA and injecting them into the perivitelline space of 48 hpf wild-type zebrafish embryos, and the images were taken with a fluorescence microscope, as described in the Materials and methods section. The number of migrated cells per zebrafish embryo was counted manually using FIJI software. Representative images from 72 h post-injection are shown. Scale bar, 100 µm and magnification, 10X. The graph represents the mean ± SD from n > 25 embryos. Statistical analysis was performed using non-parametric two-tailed Mann-Whitney U-test