Rabbit anti plp

Mouse anti-BCAS1 (Santa Cruz, 1:500, ICC and IHC, RRID: AB_10839529), mouse anti-βIII (BioLegend, 1:1000, ICC, RRID:AB_10063408), rabbit anti-βIII (BioLegend, 1:2000, RRID: AB_2564645), sheep anti-BrdU (Abcam, 1:1000, ICC, RRID:AB_302659), rabbit anti-CC3 (Millipore, 1:500, ICC, RRID: AB_91556), rabbit anti-GFAP (Dako, 1:1000, ICC, RRID: AB_10013382), rat anti-GFAP (Thermo Fisher, 1;1000, RRID:AB_2532994), chicken anti-eGFP (Abcam, 1:1000, IHC, RRID: AB_300798), mouse anti-Ki67 (BD Pharmingen, 1:300, ICC, RRID: AB_396287), rat anti-MBP (a. a. 82-87) (Millipore, 1:500, ICC, RRID: AB_94975), goat anti-PDGFRα (R&D Systems, 1:400 for IHC, 1:300 for ICC, RRID: AB_2236897), rabbit anti-PDGFRa (Santa Cruz, 1:300 for IHC, RRID: AB_631064), rabbit anti-PLP (Abcam, 1:500, ICC, RRID: AB_776593), rabbit anti-SOX2 (Cell Signalling, 1:2000, ICC, RRID: AB_2194037), goat anti-SOX2 (R&D Systems, 1:1000, IHC, RRID: AB_355110). Fluorescently labeled highly cross-absorbed secondary antibodies were purchased from Jackson ImmunoResearch and used at 1:1000 dilution. If MOM kit was used, Cy3-, DTAF-, or Cy5 conjugated streptavidin (Jackson ImmunoResearch) were used at 1:1000 dilution.

Cell sorting was carried out using a FACSAria III (BD Biosciences, San Jose, CA, USA). Dead cells were excluded using an Aqua Live/Dead staining reagent (Invitrogen, Carlsbad, CA, USA) and microglial/myeloid cells by positive staining for CD45 and CD11b. Sorted EYFP⁺ ODCs were spun onto glass slides (600 rpm) (Shandon Southern Instruments, Sewickley, PA, USA). Cells were fixed in 4% paraformaldehyde (10 min), incubated with 4% normal goat serum in 0.2% Triton-PBS, and then immunostained. For primary antibodies, goat anti-GFP antibody (Rockland, Gilbertsville, PA, USA), mouse anti-CC1 (Calbiochem, Darmstadt, Germany), rabbit anti-PLP (Abcam, Cambridge, UK), and Isolectin b4-Alexa Fluor 568 (Sigma-Aldrich, St. Louis, MO, USA) were used as indicated and were detected with secondary anti-goat Alexa Fluor 488-, anti-mouse Alexa Fluor 405-, anti-rabbit Alexa Fluor 647-conjugated antibody (all by Invitrogen, Carlsbad, CA, USA). Confocal microscopy was performed using a SP5 confocal laser microscopy (Leica, Wetzlar, Germany) and images analyzed using Imaris software (version 7.5.1; Bitplane, Zurich, Switzerland).