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. 2016 Jul 5;17(7):1072. doi: 10.3390/ijms17071072

Figure 2.

Figure 2

SSL5 binds MMPs and PSGL-1 simultaneously. (A) Formation of the triple MMP-SSL5-PSGL-1 complex was examined by far Western blot. MMP8 (0.2 µg/lane) was loaded on a gel that was subsequently blotted and incubated with 10 µg/mL SSL1 or SSL5, after which 2 µg/mL PSGL-1-Fc was added and both SSL and PSGL-1 binding was assessed using anti-HIS (anti-X-Press) and anti-Fc detection, respectively. One representative experiment is shown; (B) PSGL-1 (10 µg/mL) binding to SSL5 (3 µg/mL, coated) was not affected by the addition of (10 µg/mL) MMP8 or MMP9 in ELISA. PSGL-1 binding was assessed by anti-Fc detection and data points represent the mean absorption and SE of at least three independent experiments; (C) Addition of a 10-fold excess of PSGL-1 (10 µg/mL) to the fluorogenic peptide substrate assay did not affect the SSL5 (1 µg/mL) inhibitory potential on MMP8 (left) or MMP9 (right). Data points represent the relative mean fluorescence and SE of at least three independent experiments.