FIGURE 7.

The effects of H₂O₂ on cell migration are mediated through PML control of ITGB1 expression. A, MDA-MB-231 cells transfected with control oligonucleotide or a combination of two siRNAs against PML were subjected to wound-healing assays in the presence of control IgG or anti-ITGB1 antibodies. The results shown indicate changes in six independent fields and are representative of two independent experiments. Results for each independent siPML construct are found in supplemental Fig. S2. B, MDA-MB-231 cells were treated with H₂O₂ as described in Fig. 1C. Cells were harvested and whole cell lysates or RNA prepared. Whole cell lysates were analyzed by immunoblotting with anti-ITGB1 and anti-α-tubulin antibodies (left panel). Tubulin served as a loading control. Total RNA was analyzed by real-time RT-PCR for ITGB1 expression. GAPDH served as an internal control (right panel). C, MDA-MB-231 cells transfected with control oligonucleotide or siRNA against PML were treated with or without 100 μm H₂O₂ for 120 min as indicated. Whole cell lysates were analyzed by immunoblotting with anti-ITGB1 and anti-α-tubulin antibodies. Tubulin served as a loading control. D, MDA-MB-231-shLuc and MDA-MB-231-shPin1 cells were treated with H₂O₂ followed by whole cell lysate preparation and immunoblotting with anti-ITGB1 and anti-α-actin antibodies. E, confluent MDA-MB-231-shLuc and MDA-MB-231-shPin1 cells were treated with H₂O₂ prior to scratch wound-healing assays. Error bars indicate S.D.