Goat anti olig2

Tissue immunofluorescence staining was performed using a series of 10-µm-thick sections (Leica Instruments). Cells were cultured in a complete medium containing 10% FBS and plated on slides coated with poly-D-lysine (PDL; Sigma-Aldrich, 5 ×). After 5 days, cells were fixed with 4% paraformaldehyde for 20 min at 4 °C followed by the permeabilization of cells for 10 min with 0.1% (vol/vol) Triton X-100. Cells were then blocked with 1% BSA solution in phosphate-buffered saline (PBS). Sections and cell climbing slices were incubated with the following primary antibodies: rat anti-Nestin (1;200, Abcam, MA, USA), rabbit anti-NeuN (1:200, Abcam, MA, USA), rabbit anti-GFAP (1:300, CST, MA, USA) or goat anti-Olig2 (1:200, Abcam, MA, USA) antibodies at 4 ℃ overnight. After washing with PBS, sections were then incubated with Alexa Fluor 555-conjugated chicken anti-goat IgG (1:1000; CST, MA, USA) antibody, Alexa Fluor 555-conjugated goat anti-rabbit IgG (1:1000; CST, MA, USA) antibody, and Alexa Fluor 488-conjugated goat anti-rat IgG (1:1000; CST, MA, USA). Finally, the coverslips were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Beyotime) and imaged with a confocal microscope (Zeiss LSM800, Germany).

Immunocytochemical stainings were performed as previously described (Steffenhagen et al., 2012 (link)). The following antibodies and dilutions were used. Primary antibodies: rabbit anti-GFAP 1:1,000 (Dako, Denmark); mouse anti-myelin basic protein (anti-MBP) 1:750 (SMI-94, Covance, Anopoli Biomedical Systems, Eichgraben, Austria); rabbit anti-Galactocerebroside (GalC) 1:200 (Millipore, Burlington, MA, USA); rabbit anti Ki67 1:500 (Thermos Sc.); goat anti olig2 1:200 (Abcam); Secondary antibodies: donkey anti-mouse, rabbit, goat conjugated with Alexa Fluor 488, Alexa 568 1:1,000 (Molecular Probes, Eugene, OR, USA). Nuclear counterstaining was performed with 4′,6′-diamidino-2-phenylindole dihydrochloride hydrate at 0.25 μg/μl (DAPI; Sigma, Germany). For more details see Supplementary Materials.