Figure 3

Functional analysis of nucleofected BMP2 and BMP3-responsive promoter reporters. Human primary calvarial suture cells (1.0 x 10⁶) were nucleofected with 3 μg of BMP2 responsive promoter construct (pID183-luc; 3A) or BMP3 responsive promoter construct (p3TP-lux-Luc; 3B) using Amaxa kit-L and the T030 program. The cells were also co-nucleofected with pRL-TK-luc construct as an internal control for transfection efficiency. Each construct was incubated with BMP2 or BMP3 or vehicle as indicated in the figures. The fold induction shown is the ratio of luciferase activity from BMP2-treated or BMP3-treated to vehicle treatment (ie no BMP) at the appropriate time point. In Figure 3A, * shows responses that are significantly different to vehicle, and to BMP2 treatment at 8 hrs, and # shows that 24 hrs BMP2 treatment is significantly different to all other time-points and treatments. In Figure 3B, * shows that the response to 50 ng/ml BMP3 is significantly different compared to vehicle and to the lower BMP3 dose. The data shows a representative result from three independent experiments, each performed with triplicate biological samples.