Immunostaining Analysis of Cell Markers

Immunostaining analysis was performed as previously described (Yang et al., 2009 (link); Dincman et al., 2012 (link)). Anti-mouse A2B5 IgM and anti-mouse O4 IgM (50%, v/v) were produced by hybridoma culture. Anti-mouse Olig2 (1:1000), anti-mouse GFAP (1:1000), and anti-rabbit neurofilament (1:1000) were purchased from Millipore. Anti-rat MBP (1:500) was obtained from Abcam, anti-rabbit β-tubulin (1:1000) from Sigma, and anti-rabbit PDGFRα (1:500) from Santa Cruz. The Alexa-488 or Alexa-594 conjugated secondary antibodies were purchased from Invitrogen. The nucleic acid dye 4′,6-diamidino-2-phenylindole (DAPI) was obtained from Roche. All quantitative data are presented as means ± SD. Statistical significance of the difference was evaluated by Student’s t-test. P < 0.05 was considered statistically significant.