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. 2021 Oct 21;297(5):101323. doi: 10.1016/j.jbc.2021.101323

Figure 6.

Figure 6

ADAMTS8 cleaves human OPN.A, OPN (760 nM) was incubated with either ADAMTS8 WT or EQ (50–500 nM) for 24 h at 37 °C. Samples were subjected to 4 to 12% SDS-PAGE/immunoblot under reducing conditions and probed with a polyclonal anti-OPN antibody. B, inhibition of ADAMTS8 proteolytic activity. TIMP-1, -2, -3, and -4 (each at 1 μM) were incubated with ADAMTS8 (500 nM) for 1 h at 37 °C before addition of OPN. Following SDS-PAGE and immunoblotting, OPN fragments were detected by a polyclonal anti-OPN antibody. A representative immunoblot is shown (n = 4). Full-length (FL) substrates and specific cleavage fragments (CF) are indicated. C, densitometric analysis showing quantification of ADAMTS8-generated cleavage fragments (CF). Data are presented as mean ± SEM (n = 4). ∗p < 0.05 relative to buffer control, without inhibitor (Mann–Whitney test).