Figure 5.

Knockdown of RUNX2 enhances ADR-mediated apoptosis. 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. Twelve hours after ADR exposure, images were taken (a) and floating and attached cells were collected, fixed in 70% ethanol, stained with propidium iodide (PI) and subjected to FACS analysis (b). Cells with sub-G1 DNA content were considered dead. Under the same experimental conditions, cell lysates and total RNA were prepared and analyzed by immunoblotting (c) and RT-PCR (d), respectively. Actin and GAPDH were a loading control for immunoblotting and an internal control for RT-PCR, respectively