Figure 4.

BMP9-cm effectively induces the expression of downstream target genes in iMEFs. Subconfluent iMEFs were stimulated with BMP9-cm or Con-cm (at 25% concentrations), or infected with AdBMP9 or AdGFP (MOI=10), and maintained in 1% FBS DMEM. At the indicated time points, total RNA was isolated using TRIzol reagents, subjected to reverse transcription reactions, and quantitative real-time PCR (qPCR) analysis using primers specific for mouse Smad6, Smad7, Id1, Id2, Id3, and CCN1 transcripts. Fold expression was calculated by dividing BMP9-induced gene expression with the gene expression level in Con-cm or AdGFP groups. The qPCR reactions were done in triplicate.