Olig 1

Cells or cryosectioned tissues (10 μm) were post-fixed with 4% paraformaldehyde at room temperature for 10 min. After permeabilization with PBS containing 0.5% Triton and 2% goat serum (blocking solution), the primary antibodies were incubated overnight at 4 °C. Cells or tissue sections were incubated with GAP43 (α-rabbit; Cat. ab128005 Abcam, Hong Kong, China), β-tubulin III (α-mouse; Cat. MO15052 Neuromics, Edina, MN, USA), GFAP (α-rabbit; Cat. Z0334 DAKO, Santa Clara, CA, USA), RIP (α-mouse; Cat. MAB1580 Chemicon, Pittsburgh, PA, USA) and OLIG1 (α-rabbit; Cat. AB5320 Chemicon). Primary antibodies were diluted 1:200 in blocking solution. After being rinsed three times with PBS, the cells or the tissue sections were incubated with Oregon Green-Alexa488, Alexa555 or Alexa647 dye-conjugated secondary antibodies for 1 h at room temperature. All cells and tissue sections were counterstained by incubation with DAPI for 5 min at room temperature followed by washing steps. Signals were visualized by both fluorescent microscopy (fluorescence microscope Leica DM6000B, Wetzlar, Germany) and confocal microscopy (confocal microscope Leica TCS-SP2-AOBS, Wetzlar, Germany). The quantification of immunostainings was performed using ImageJ software (Image J2, Madison, WI, USA).