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. 2017 Nov 30;3(2):e000604. doi: 10.1136/rmdopen-2017-000604

Figure 2.

Figure 2

Suramin increases TIMP3 levels and reduces MMP and aggrecanase activity in vitro. (A) Western blot showing an increased amount of TIMP3 protein in extracellular matrix (rest fraction) of suramin-treated ATDC5 micromasses compared with controls after 21 days of differentiation culture. (B) Immunohistochemistry showing increased TIMP3 and decreased VDIPEN and NITEGE levels in suramin-treated micromasses compared with controls after 21 days of differentiation culture. Scale bar 50 µm. (C) Fluorescence intensity was measured on extracellular matrix proteins from micromasses (rest fraction) treated with suramin or vehicle, after anti-TIMP3 immunoprecipitation. Values are shown as the difference with the fluorescence emitted from anti-IgG immunoprecipitates. (D) Ten-micromole TIMP3 neutralising antibody blocks the effects of suramin in the ATDC5 micromasses at day 14; control is 10 µM IgG Ab. (*P<0.01, one-way analysis of variance followed by Tukey’s multiple comparisons test). Data shown are representative of three (A, B) or two (C) biological replicates. Error bars indicate mean±SEM of three technical replicates per experiment.