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. 2016 Feb 24;7(13):16462–16478. doi: 10.18632/oncotarget.7656

Figure 4. miR-30a decreases the invasiveness of breast cancer cells.

Figure 4

(A) MDA-MB-231 cells were transfected with plemiR or plemiR-30 and then treated with miR-30a inhibitor (anti-miR-30a) or underwent Slug knockdown with a Slug-specific short hairpin RNA (sh-Slug) or sh-Luc, a control shRNA. The cells were plated in modified Boyden chambers with polycarbonate membranes containing Matrigel and cultured for 12 h. Cells were then fixed, stained with Giemsa solution, and photographed (× 200). Upper panel: Cells that invaded through the pores onto the lower side of the filter. Lower panel: The invading cells were counted in eight randomly chosen microscope fields. Data are shown as the mean ± SD from three independent experiments. *P < 0.05, **P < 0.01. (B) Reduced Slug and fascin expression, but increased claudin levels, as determined by western blot analysis, in MDA-MB-231 cells expressing plemiR-30a or shRNA-Slug, as compared with controls plemiR or shRNA-Luc, respectively. In addition, Slug and fascin protein levels were restored in plemiR-30a–expressing/MDA-MB-231 cells transfected with anti-miR-30a. β-actin was the loading control. (C) Knockdown of Slug notably decreases the invasion of MDA-MB-231 cells, whereas overexpression of miR-30a fails to abrogate the reduced frequency of invasion for sh-Slug/MDA-MB-231 cells. (D) The expression of Slug, fascin, and claudins was also analyzed by western blotting. β-actin was the loading control.