Fig. 3

Bioactivity of purified recombinant Shh proteins. a Upper panel represents the schematic diagram of plasmid p8XGBE-Luc. Lower panel shows that Shh increases Gli-mediated reporter activity. NIH3T3 cells transfected with p8XGBE-luc and pRSV-βGAL were stimulated during 18 h with 0.5, 1.0, 5.0, 10, and 100 μg/ml of purified Shh. Bars represent the mean SEM of the firefly to β-galactosidase activity ratio of three independent experiments and are expressed as the fold with respect to no treatment. b C₂C₁₂ cells were cultured in the presence of BMP-4 (50 ng/ml), Shh-N (500 ng/ml), or both for 72 h. CTRL cells were not treated with any proteins. AP activity was measured in cell lysates and normalized to total protein concentration. *p < 0.05 and **p < 0.005 vs. untreated. Error bars represent standard deviations