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. 2014 Dec 11;14(1):67–77. doi: 10.1111/acel.12298

Figure 5.

Figure 5

GzmB-mediated decorin cleavage renders collagen fibrils more susceptible to degradation by MMP-1. (A) Decorin cleavage assay. Recombinant decorin (0.4 μg) was treated with GzmB (100 nm) ± Inhibitor (I; 50 μm) for 8 h, 37°C. (B) Dorsal skin sections from control and UV-irradiated mice were stained with decorin antibody. Intensity of staining in the dermis (excluding hair follicles) was measured by detecting the number of positive pixels above a set threshold, normalized to area. Results are expressed as a percentage of wild-type control (WT-C) (mean ± SEM; *P < 0.05; ***P < 0.001 Tukey's multiple comparison). Scale bars = 60 μm. (C) Collagen fibrils ± decorin were treated with or without GzmB (100 nm) ± Compound 20 (I, 50 μm) as indicated. Loss of intact collagen (closed arrowhead) was assessed by SDS-PAGE electrophoresis followed by Coomassie blue staining. Open arrowhead = bovine serum albumin. Results are expressed as a percentage of intact collagen control (mean ± SEM of three independent experiments, *P < 0.05; ***P < 0.001 Tukey's multiple comparison).