Figure 5

Simultaneous analysis of multiple BMPs in a serumsample. C2C12BRE cells were treated as in Figure 4 with increasing concentrations of BMP4 (A), BMP6 (B) or BMP9 (C) within the linear range. Standard curves for (A) BMP4, (B) BMP6 and (C) BMP9 were calculated. (D) Other C2C12BRE cells were incubated with 10% FCS, from Autogen Bioclear, batch 212-181107, in the absence or presence of 5 ng/ml BMP4 antibody, 2 ng/ml BMP6 antibody or 0.1 ng/ml BMP9 antibody. After 16 hours of treatment, cells were lysed and luciferase activity and protein content were determined and the relative luciferase units were calculated as described in Figure 2. (E) Δ luciferase units for each BMP were calculated as described in Figure 4, and the BMP content was calculated by extrapolating the Δ luciferase units' value for each BMP in the appropriate standard curve.