Anti olig1

The sections used for immunofluorescence staining were incubated with primary monoclonal mouse anti-MBP antibody (1:500, Abcam, Cambridge, MA), anti-GAP43 and anti-CD68/ED1 (1:100; Santa Cruz, Dallas, TX) antibodies and polyclonal rabbit anti-activated caspase-3 (1:500; Cayman Chemical, Ann Arbor, MI), anti-NF200 (1:500, Abcam, Cambridge, MA), anti-GFAP (1:200, Thermo Fisher Scientific Waltham, MA), anti-Olig1 (1:500; Abcam, Cambridge, MA), anti-CNTF (1:500, Abcam, Cambridge, MA), anti-NF-kB p65 (1:500, Abcam, Cambridge, MA) antibodies and anti-BDNF(1:500, Abcam, Cambridge, MA) antibodies overnight at 4 °C. The sections were then washed with phosphate-buffered saline (PBS) and incubated with 1:200 TRITC (rhodamine)-conjugated secondary antibodies for 1 h at 37 °C (Invitrogen, Carlsbad, CA). The sections were finally coverslipped with Antifade Gel/Mount Aqueous Mounting Media (Southern Biotech, Birmingham, AL).
Five sections from the brain cortex and anterior horns of the spinal cord for each animal were randomly selected and images were photographed under 200x magnification in three vision fields per section. GFAP, MBP, CD68-immunoreactive areas were analyzed with NIH image software, and the numbers of GAP43, caspase-3, NF-200, Olig1, CNTF, NF-kB p65, BDNF-labeled cells were counted.

The following primary antibodies were used: anti-MBP (rat, 1:800, Chemicon),
anti-neurofilament (mouse, 1:800, SMI31, Covance), anti-Olig1 (rabbit, 1:1000,
Abcam), anti-p75^NTR (rabbit, 1:500, Promega), anti-S100 (rabbit,
1:500, Dako).
The following secondary antibodies were used: donkey-anti-rabbit AlexaFluor488,
donkey-anti-mouse DyLight549, donkey-anti-rat AlexaFluor647 (all 1:500, Jackson
ImmunoResearch Laboratories), DAPI (4′,6-diamidino-2-phenylindole)
(1.25 μg/ml; Molecular Probes) was used as cellular counter
stain.

Sections of brain tissue were deparaffinized in graded alcohol solutions and xylene. The sections were then blocked with 3% H₂O₂ (37°C, 30 min) and goat serum (37°C, 20 min), followed by addition of rabbit anti-rat monoclonal anti-OLIG1 (1:300 dilution; Abcam, San Francisco, CA, USA) and incubation overnight at 4°C. The negative control tissues were incubated with phosphate-buffered saline (PBS) instead of the primary antibody, and also incubated with the biotin-labeled goat anti-rabbit IgG secondary antibody (37°C, 30 min), similarly to the tissues of interest. All sections were then incubated with a horseradish peroxidase (HRP) marker (37°C, 30 min), stained with 3′-diaminobenzidine (DAB), re-stained with hematoxylin, dehydrated, vitrified, and mounted. Ten different sections from each time-point were randomly selected from each group, and the periventricular alba was observed in 5 random fields of a light microscope (magnification, ×400).