Anti sulf2

The following antibodies were used in this study: anti-SULF2, anti-IL-6, and anti-β-catenin from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-Bcl-X_L, anti-STAT3, and anti-phospho-STAT3 from Cell signaling Technology (Danvers, MA, USA); anti-β-actin from Sigma-Aldrich (St. Louis, MO, USA). Recombinant human IL-6 was purchased from Millipore (Bedford, MA, USA).

Cryostat (10-μm thick) sections of paraformaldehyde (PFA)-fixed brains were used for immunohistochemistry. To detect Sulf or Slit proteins and HS, fresh-frozen sections fixed with 95% ethanol and 1% acetic acid were used^{42 (link)}. The primary antibodies used were anti-neurofilament-M (1:1000; Zymed), anti-FLAG (1:500; Abcam, ab6711), anti-GFP (1:1000; Molecular Probes), anti-Sulf1 (1:50; Abcam, ab32763), anti-Sulf2 (1:50; Santa Cruz, M79), anti-HS (1:5; AO4B08; refs^{18 (link),19 (link)}), anti-nestin (1:1000; Chemicon), and anti-laminin (1:1000; Sigma). The secondary antibodies used were peroxidase-conjugated anti-mouse IgG (1:200; Chemicon) and Alexa488- or Alexa568-conjugated anti-rabbit IgG (1:250; Molecular Probes). A chromogenic reaction was carried out using a VECTASTAIN Elite ABC kit and a DAB substrate kit (Vector Laboratories). For immunofluorescence, cell nuclei were stained with 3.3 μM TO-PRO-3 iodide (Molecular Probes). Whole-mount immunostaining was done (with minor modifications) using the same reagents after the meninges were removed^{43 (link)}.