NSCs were prepared from embryonic day 14.5 mouse telencephalon and expanded in serum-free Dulbecco's Modified Eagle's medium (DMEM) /F12 medium containing bovine insulin (10 μg/ml), human transferrin (100 μg/ml), BSA (100 μg/ml), progesterone (60 ng/ml), putrescine (16 μg/ml), sodium selenite (40 ng/ml), N-acetylcysteine (60 μg/ml), forskolin (5 μM), PDGF (10 ng/ml), penicillin, streptomycin, and bFGF as floating spheres. Astrocytes were induced in NSC cultures by changing the medium to DMEM with 10% FCS for 3 wks. OPCs were induced by culturing NSCs in DMEM containing PDGFAA (10 ng/ml), bFGF (2 ng/ml), and 0.25% FCS and purified by sequential immunopanning.
Extracted molecule
total RNA
Extraction protocol
Total RNA was prepared using a RNeasy Mini Kit (QIAGEN).
Label
biotin
Label protocol
Five micrograms of total RNA were labeled with biotin by in vitro transcription (IVT) using the One-Cycle Target Labeling procedure for biotin labeling by in vitro transcription (Affymetrix), according to the manufacturer’s instruction.
Hybridization protocol
Labeled probes were used for the GeneChip Mouse Genome 430 2.0 array (Affymetrix) according to the manufacturer’s instructions.
Scan protocol
The microarray image data were processed with the GeneChip Scanner 3000 (Affymetrix) to generate CEL data.
Data processing
The CEL data were then analyzed with dChip software, which can normalize and process multiple datasets simultaneously.
