Mouse brains were processed and immunostained as previously described [9 (link)–12 (link)]. In brief, mice were decapitated under isoflurane anesthesia and brains were harvested and drop fixed in 4% formaldehyde for 24 h, then transferred to graded ethanol series, embedded in paraffin, and sectioned. Eyes were processed and fixed as previously described [13 (link)]. In brief, eyes were harvested after decapitation, injected with 4% formaldehyde, and then drop fixed, embedded in paraffin, and sectioned.
Primary antibodies used were: BAK diluted 1:200 (Cell Signaling, #12105), Myelin Basic Protein (MBP) diluted 1:1000 (Abcam, #ab7349), SOX10 diluted 1:100 (Santa Cruz, #sc-17342), cleaved-Caspase 3 (cC3) diluted 1:400 (Biocare Medical, #CP229C), glial fibrillary acidic protein (GFAP) diluted 1:2000 (Dako, Z0334), PDGFRA diluted 1:200 (Cell Signaling, #3174), SOX2 diluted 1:200 (Cell Signaling, #4900S), SOX9 diluted 1:200 (R&D Systems, #AF3075), NESTIN diluted 1:500 (Cell Signaling, #4760), and IBA1 diluted 1:2000 (Wako Chemicals, #019-19741). Stained images were counterstained with DAPI, digitally imaged using an Aperio Scan Scope XT (Aperio), and subjected to automated cell counting using Tissue Studio (Definiens).
Primary antibodies used were: BAK diluted 1:200 (Cell Signaling, #12105), Myelin Basic Protein (MBP) diluted 1:1000 (Abcam, #ab7349), SOX10 diluted 1:100 (Santa Cruz, #sc-17342), cleaved-Caspase 3 (cC3) diluted 1:400 (Biocare Medical, #CP229C), glial fibrillary acidic protein (GFAP) diluted 1:2000 (Dako, Z0334), PDGFRA diluted 1:200 (Cell Signaling, #3174), SOX2 diluted 1:200 (Cell Signaling, #4900S), SOX9 diluted 1:200 (R&D Systems, #AF3075), NESTIN diluted 1:500 (Cell Signaling, #4760), and IBA1 diluted 1:2000 (Wako Chemicals, #019-19741). Stained images were counterstained with DAPI, digitally imaged using an Aperio Scan Scope XT (Aperio), and subjected to automated cell counting using Tissue Studio (Definiens).
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