Mab1326

Cells were fixed with 4% PFA in phosphate buffered saline (PBS) for 15 min at room temperature. After washed with PBS three times, the cells were blocked and permeated with PBS containing 1% BSA and 0.1% Triton (without Triton for O4 staining) for 30 min at room temperature. Then cells were incubated with the relevant primary antibody at 4°C overnight. After PBS washing for three times, cells were incubated with the appropriate fluorescence‐conjugated secondary antibody for 1 hr at room temperature. Nuclei were stained with Hoechst 33,342 (10 mg mL⁻¹). Antibodies used in this assay are as follows: anti‐Nestin (Millipore, MAB353, 1:400), anti‐A2B5 (Sigma, A8229, 1:500), anti‐Olig2 (Millipore, AB9610, 1:200), anti‐MBP (Covance, SMI‐94R, 1:500), anti‐Ki67 (Cell signaling, 9129S, 1:300), anti‐MBP (Abcam, ab7349, 1:750), anti‐O4 (R&D Systems, MAB1326, 1:1,000), anti‐CNPase (Millipore, MAB326, 1:500), anti‐NF‐200 (Sigma, N4142 and N5389, 1:1,000), anti‐Caspr (Abcam, ab34151, 1:500). TUNEL staining was carried out according to the manufacturer's instructions (Roche, 11684795910).

Cells were fixed with 4% PFA in phosphate‐buffered saline (PBS) for 15 min at room temperature and blocked in PBS containing 1% BSA and 0.3% Triton for 30 min at room temperature. Then cells were incubated with the relevant primary antibody at 4°C overnight and the appropriate fluorescence‐conjugated secondary antibody for 1 hr at room temperature. Nuclei were stained with Hoechst 33,342 (10 mg/ml). Antibodies used in this assay were as follows: anti‐MBP (Covance, SMI‐94R, 1:500), anti‐NF‐200 (Sigma, N4142, 1:1000), anti‐O4 (R&D Systems, MAB1326, 1:1200), and anti‐CNPase (Millipore, MAB326, 1:1200).