Figure 2.

Cleavage of LOX by ADAMTS2 and ADAMTS14. LOX cleavage was assayed by immunoblotting in LOX/ADAMTS-overexpressing co-cultures after incubation with doxycycline for 48 h (A and B) and after in vitro incubation assays (C and D) for the indicated times. For comparison, LOX fragments generated upon incubation with BMP1 are also shown in C. ADAMTS2 and ADAMTS14, either in co-culture or using purified proteins, promoted the accumulation of a mature form of about 25 kDa (red arrow), whereas BMP1 cleavage gave rise to bands in the 30-kDa range (blue arrow), indicating distinct processing sites. ADAMTS3 co-culture did not modify the relative levels of both fragments, suggesting it was not able to process LOX under our experimental conditions. E, sequential incubation with BMP1 and ADAMTS14. LOX supernatant was first incubated with BMP1 for 1 h. Then, ADAMTS14 was added into the reaction mixture for the indicated times.