Antibodies and Reagents

Immunoblot analysis of astrocyte cell lysates was performed using antibodies, including anti-β₃ integrin (AB2984; Millipore), anti-Δ-heparan sulfate (3G10; Seikagaku), anti β actin (A5060; Sigma-Aldrich), and secondary horseradish peroxidase-conjugated goat anti-mouse (074-1806) or anti-rabbit (074-1516) IgG polyclonal secondary antibodies from KPL. Anti-Syndecan-4 (sc-12766; Santa Cruz) was used to detect the recombinant Syndecan-4-Fc. Goat polyclonal antibody anti-rat β₃ integrin (sc-6627; Santa Cruz) and rabbit polyclonal anti-rat β₁ integrin (sc-8978; Santa Cruz) were used as blocking antibodies in the co-culture assays. Reagents used in this work were: Heparin (H-3400; Sigma-Aldrich), Heparitinase III from Flavobacterium heparinum EC 4.2.2.8 (H-8891; Sigma-Aldrich), Cell TrackerTM Green 5-chloromethyl fluorescein diacetate dye (C2925; Invitrogen) for monitoring living cells. The cell Line Nucleofector Kit (VCA-1003; Lonza) was used to transfect the astrocytes in the Nucleofector Device, X-treme GENE HP DNA Transfection Reagent (06-366 244 01; Roche) for HEK293T cell transfection, 1-Step Ultra TMB (3,3′,5,5′-tetramethylbenzidine) ELISA substrate (34028; Thermo Scientific), recombinant human bFGF protein (PH-G0266; Gibco), bovine serum albumin fraction V protease- and immunoglobulin-free (BSA-50; Rockland), protein A-sepharose from Staphylococcus aureus (P-3391; Sigma) and protein G-polystyrene beads (3.1 and 2.1 μm; Spherotech).