Figure 7.

siRNA-mediated knockdown of RUNX2 enhances ADR-dependent cell death and expression of TAp73. (A) Knockdown of RUNX2. U2OS cells were transfected with control siRNA or with siRNA against RUNX2. Twenty-four hours after transfection, cells were treated with 0.5 μm of ADR or left untreated. After 24 h of incubation, cells were stained with anti-RUNX2 antibody (green). Cell nuclei were stained with DAPI (blue). Scale bar = 25 μm. (B) Genomic DNA fragmentation. U2OS cells were transfected as described in (A). Twenty-four hours post-transfection, cells were exposed to 0.5 μm of ADR or left untreated. Twenty-four hours after treatment, attached and floating cells were collected, and their genomic DNA was prepared and analyzed by 1% agarose gel electrophoresis. (C) RT-PCR analysis of the endogenous TAp73. U2OS cells were transfected as described in (A). Transfected cells were treated with 0.5 μm of ADR or left untreated. Twenty-four hours after treatment, total RNA was prepared and subjected to RT-PCR analysis.