Aggrecan 6 b 4

The following primary Abs were used in this study: Aggrecan (6-B-4) (Abcam, Cambridge, MA, USA), ARGXX (BC-3) (Abcam), phospho-JNK1/2 (Thermo Fisher Scientific, Waltham, MA, USA) β-actin (Cell Signaling Technology, Danvers, MA, USA), FLAG-M2 (MilliporeSigma, Burlington, MA, USA), acetylated α-tubulin (6-11B-1) (MilliporeSigma), early endosome antigen 1 (EEA-1; Abcam), LRP-1 β-light chain (Abcam), LRP-1 α-heavy chain (8G1) (Abcam), a disintegrin and metalloproteinase (ADAM)-17 (Abcam), MMP-13 (Santa Cruz Biotechnology, Dallas, TX, USA), MMP14 (anti-catalytic domain EP1264Y) (Abcam), mouse monoclonal anti–MT1-MMP hemopexin domain 222-1D8 (a gift from Yoshi Itoh, Kennedy Institute) generated as previously described (41 (link)), and arl13b (ProteinTech, Rosemont, IL, USA). Anti-AGEG was a kind gift from Hideaki Nagase (University of Oxford). Purified aggrecan from bovine cartilage came from MilliporeSigma. Recombinant human C-terminal His-tagged human receptor-associated protein (RAP) was produced in Escherichia coli using a pET3a-based expression vector and purified as described previously (13 (link)). The domain deletion mutant, ADAMTS-5 (TS5-3)-flag, is described in Gendron et al. (42 (link)), and recombinant MMP-13 is described in Yamamoto et al. (43 (link)). Recombinant IL-1β was purchased from PeproTech (London, United Kingdom).