Antibodies (Abs) used were: Anti-α2δ-1 Ab (mouse monoclonal, Sigma-Aldrich), anti-α2δ-3 and anti-δ-3 Ab,8 (link) anti-HA Ab (rat monoclonal, Roche), anti-GAPDH Ab (mouse monoclonal, Ambion), anti-FLAG Ab (rabbit polyclonal; Sigma), anti-PDI (mouse monoclonal, Ambion), anti-g97 (rabbit polyclonal; Abcam), and anti-flotillin Ab (monoclonal, BD Biosciences). For immunoblotting, secondary Abs (1:2000) were anti-rabbit–Horseradish Peroxidase (HRP), and anti-mouse HRP (Biorad). For immunocytochemistry, anti-rat-Alexa Fluor 594 was used at 1/500 (ThermoFisher).
The metalloprotease inhibitors GM6001 (BML-EI300, Enzo Life Sciences), SB-3CT (BMEI325, Enzo Life Sciences), and MMP-13 inhibitor (BML-EI302, Enzo Life Sciences) were dissolved in DMSO (or water for MMP-13 inhibitor) and used at the concentrations stated. N-TIMP-3 protein (expressed in Escherichia coli as previously described,38 (link)) or control samples in the absence of N-TIMP-3, were preincubated with heparin (200 µg/ml) for an hour at 37°C before adding to the cells.
The metalloprotease inhibitors GM6001 (BML-EI300, Enzo Life Sciences), SB-3CT (BMEI325, Enzo Life Sciences), and MMP-13 inhibitor (BML-EI302, Enzo Life Sciences) were dissolved in DMSO (or water for MMP-13 inhibitor) and used at the concentrations stated. N-TIMP-3 protein (expressed in Escherichia coli as previously described,38 (link)) or control samples in the absence of N-TIMP-3, were preincubated with heparin (200 µg/ml) for an hour at 37°C before adding to the cells.