Fig. 5.

Bioactivity of Tsg for BMP-4 and-7. Inhibition of BMP-4 and BMP-7 growth factor activity by Tsg was detected by inhibition of alkaline phosphatase production in C2C12 cells (A and B) and Smad-1/− 5 phosphorylation in HEK293T cells (C and D) with a 28 × molar excess of Tsg. A rapid qPCR assay based on the Id3 expression in C2C12 cells after 5 h of incubation with growth factor was also used. A range of Tsg concentrations were tested with BMP-4 (E) and BMP-7 growth factor (F). (G) Natively deglycosylated Tsg was still able to inhibit BMP-7 as detected by inhibition of alkaline phosphatase production. Errors represent standard deviation from at least three independent experiments.