FIGURE 4.

RUNX2 regulates genes that play a role in cell motility. RUNX2 knockdown with two different siRunx2 oligonucleotides (48 h) in SAOS-2, U2OS, and MDA-MB-231 cells affects several genes that are involved in motility and adhesion of cells as identified with ChIP-on-chip. The bar graphs (middle panels) show average effects of the two different siRNA oligos compared with two negative nonsilencing RNAs on gene expression as measured by RT-qPCR. The errors in technical replicates within the same biological sample were negligible and not indicated. Black and gray bars indicate effects of more than 30%; white bars indicate no significant effects; Ø indicates no expression. RUNX2 knockdown at the protein level (left panels) is shown by Western blot analysis using lysates from SAOS-2, U2OS, and MDA-MB-231 cells treated with or without siRUNX2. Western blots were quantified using ImageJ (imagej.nih.gov) and revealed a >70% reduction in RUNX2 levels in all cell lines.