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. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: J Immunol. 2013 Jun 24;191(3):1404–1412. doi: 10.4049/jimmunol.1300856

FIGURE 1.

FIGURE 1

Induction of aggrecanolysis in cartilage by hTryptase-β. A-D, Measurement of aggrecan in sections of femoral head explants using toluidine blue staining (A, B) and immunohistochemistry with anti-aggrecan antibodies (C, D) showed appreciably greater aggrecan loss from cartilage when explants were incubated for 24 h with 15 μg/ml rhTryptase-β (B, D) compared to culture media alone (A, C). Isotype controls for immunohistochemistry are inset in the panels; scale bars = 50 μm. E, rhTryptase-β (15 μg/ml) induced significant release of aggrecan-derived GAGs from both live and dead femoral head explants compared to media alone. ** p < 0.01; ***, p < 0.001; error bars represent S.E.M.; n = 3. F, rhTryptase-β (12 μg/ml, lane 2) and pro-MMP-3 (50 ng, lane 3) were unable to directly degrade purified pig aggrecan (200 μg/ml) when incubated for 16 h, compared to incubation of aggrecan with media alone (lane 1), or with 50 ng pro-MMP-3 activated with 1 mM APMA (lane 4). Samples were analyzed by immunoblotting using the 2B6 antibody that recognizes the chondroitinase ABC-generated chondroitin sulfate stubs.