Fig. 5
Re-expression of DIRAS3 reversed Sp1 knockdown-mediated breast cancer cell migration and invasion. a Protein levels of Sp1 and DIRAS3 in MDA-MB-231 cells were assessed by Western blot analysis after plasmid and/or siRNA transfection. Transfection of siSp1 led to decrease in Sp1 expression as well as DIRAS3 expression. Additionally, transfection of DIRAS3 plasmid into Sp1-silenced MDA-MB-231 cells efficiently elevated DIRAS3 expression. GAPDH was used as a loading control. b MDA-MB-231 cells with distinct treatments were subjected to wound-healing assay. Representative photographs at 24 h were taken (left panel). Magnification: ×100. Percentage wound recovery was also shown (right panel). *P < 0.05. c MDA-MB-231 cells with distinct treatments were subjected to transwell migration and invasion assays. 12 h post culture, cells were stained with crystal violet. Representative photographs from each group were taken for subsequent cell number counting. Magnification: ×200. d Migratory and invasive MDA-MB-231 cells were counted and shown. Cell numbers represented average values from three microscopic visions of each group. *P < 0.05. e, f In MCF-7 cells, re-expression of DIRAS3 was also observed to reverse Sp1 knockdown-mediated breast cancer cell migration and invasion, as revealed by the crystal violet staining (e) and transmigrated cell number counting (f). Magnification: × 200. *P < 0.05. siNC, negative control siRNA