FIG. 8.

Effect of Cbfa1 and TGF-β on collagenase 3 expression in MG-63 osteosarcoma cells. Cells were transiently transfected with pcDNA3 (lanes 1 to 4) or pCMV-Osf2/Cbfa1 (lanes 5 to 8). Transfected cells were stimulated with vehicle alone, IL-1β (5 ng/ml), bFGF (5 ng/ml), or TGF-β (5 ng/ml) for 24 h. RNA from the stimulated transfected cells was further prepared and used for RT-PCR as described in Materials and Methods. Aliquots of samples were taken at 26 and 29 cycles of amplification. Samples were separated in agarose gel, transferred to nylon filters, and hybridized with collagenase 3 and actin probes. The results shown are from a representative experiment and were consistently reproducible in several independent experiments.