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. 2005 Aug 25;115(9):2373–2381. doi: 10.1172/JCI25118

Figure 4.

Figure 4

Characterization of ALP induction by GDF5 mutants in chondrogenic ATDC5 cells. ALP activity was measured after stimulation of ATDC5 cells with increasing amounts of recombinant proteins after 3 days. (A) Recombinant BMP2, GDF5, and R438L display characteristic dose-dependent inductions of ALP activity in ATDC5 cells. The R438L mutant induces significantly higher ALP activity than WT GDF5, whereas L441P is almost inactive. (B) Stimulation of ATDC5 cells with L441P or a combination of GDF5 and L441P. Cells were incubated with increasing amounts of L441P with or without 10 nM GDF5. No stimulating or suppressing effect of increasing amounts of L441P on GDF5-dependent induction of ALP activity was observed. (C) Inhibition of ALP induction by recombinant NOG in stimulated ATDC5 cells. ALP was determined after incubation with 5 nM recombinant BMP2, GDF5, R438L, or L441P, and cotreatment with increasing amounts of NOG. GDF5, R438L, and L441P display similar inhibition profiles by NOG.