FACS-purification of LeX+NG2−/EGFPneg (NSCs) in the CNP-EGFP mouse or GFAP-GFP+LeX+NG2neg (NSCs) in the GFAP-GFP mouse has been previously described19. Tissue from P8 CNP-hEGFR, CNP-EGFP/CNP-hEGFR, GFAP-GFP or CNP-hEGFR/GFAP-GFP mice was dissociated into single cell suspensions, followed by immunostaining for NG2 and LeX (MMA clone)19. SVZ neural stem cells and NPCs (after EGF or saline infusion, or 5–7days after focal demyelination of the corpus callosum) were FACS-purified using anti-LeX antibodies to purified LeX+/GFAP-GFP+ (NSCs) or GFAP-GFPnegLeX+ (NPCs) cells from the hGFAP-GFP mouse. Tissue was dissociated into single cell suspensions, followed by immunostaining for NG2 and LeX (MMA clone)19. To FACS-purify NSCs, antibodies were used in combination with appropriate R-PE- and PE-Cy5.5-conjugated secondary antibodies (Caltag, Burlingame, CA). Cell suspensions were analyzed for light forward and side scatter using a FACSAria instrument (BD, Biosciences, Franklin Lakes, NJ).
