Rabbit psmad1 5 8

IHC was performed on 16μm serial sections with blocking and antibody incubation carried out overnight at 4°C in PBS containing 0.3% Triton/10% normal goat serum with the exception of GAD67 for which Triton was excluded. For human brains, the cryosections were heated for antigen retrieval in sodium citrate buffer before blocking. Primary antibodies were used at the following dilutions: rabbit Olig2 (1:2000; Dr. Charles D Stiles, Dana Farber Cancer Institute; Boston MA), rabbit Olig1 (1:1000; Dr. Charles D Stiles, Dana Farber Cancer Institute; Boston MA), mouse MBP (1:1000; Covance), rabbit DCX (1:500; Cell Signaling); mouse BrdU (1:100; BD Biosciences); mouse GAD67 (1:500; Millipore); mouse Tuj1 (1:1000; Covance); rabbit GFAP (1:1000; Dako); rat PDGFRα (1:500 BD Biosciences), rabbit pSMAD1/5/8 (1:1000; Cell Signaling). BrdU detection was carried out as previously described^{18 (link)}, with IHC for the protein of interest carried out as described above before the application of the BrdU antibody. After primary antibody incubation and washes, sections were incubated with Alexa-fluor conjugated species directed secondary antibodies (Invitrogen) and then rinsed before being mounted with DAPI Fluoromount-G.