Effect of Notch on Hey1, Hey2 (lower panel) and Ctgf (upper left panel) mRNA, Ctgf hnRNA (upper middle panel) and Ctgf protein (upper right panel) expression in osteoblasts. Calvarial osteoblasts isolated from RosaNotch mice were transduced with Ad-CMV-Cre, to activate Notch (black bars), or control Ad-CMV-GFP (white bars) and cultured to confluence (day 0) or up to 2 weeks following confluence. Samples for hnRNA and protein determination were obtained 3 days post-confluence. Total RNA was extracted, reversed transcribed and amplified by qRT-PCR. Data for mRNA and hnRNA are expressed as Ctgf, Hey1 and Hey2 copy number corrected for Rpl38 expression relative to the mRNA or to the Ctgf hnRNA expression in Ad-CMV-GFP control cells, arbitrarily set at a value of 1. Data for Ctgf protein, measured by ELISA, are expressed as ng/ml of culture medium. Data for Ctgf, Hey1 and Hey2 mRNA were pooled from 2 experiments. Values are means ± SEM; n = 4. *Significantly different between Ad-CMV-Cre Notch activated cells and control, p < 0.05.