Origin of sample: mouse embryos expressing green fluorescent protein (GFP) in neural crest stem cells and later glial cells under the control of the proteolipid protein gene (Plp) promoter. At P0 (day of birth) the sciatic nerve was dissected out. GFP-expressing cells were isolated by FACS sorting. See also Buchstaller_suplTab1.xls in publication. RNA extracted with RNeasy kits (Qiagen). Target preparation by two rounds of in vitro transcription (Hoffmann, R., Seidl, T., Neeb, M., Rolink, A. & Melchers, F. Changes in gene expression profiles in developing B cells of murine bone marrow. Genome Res 12, 98-111 (2002)). Labeling protocol: Standard Affymetrix biotin labeling protocol (Affymetrix, Expression Analysis: Technical manual, 2000) in 2nd round of in vitro transcription. See also Buchstaller_suplTab1.xls for yields of cRNA. Hybridization procedure: Standard Affymetrix protocols (Affymetrix, Expression analysis: Technical manual, 2000). Scanning with Affymetrix scanner and MAS 5.0 software. The original images (.dat files) and .cel files are available on request (mantei@cell.biol.ethz.ch) The .cel files produced by MAS 5.0 were normalized using the program dChip (smoothing spline normalization; Li, C. & Wong, W.H. Model-based analysis of oligonucleotide arrays: model validation, design issues and standard error application. Genome Biol. 2, research0032.0031-0032.0011 (2001); http://www.dchip.org). dChip was also used to calculate a probe sensitivity index for each probe on the basis of 18 arrays, and then a "model-based expression index" (MBEI) for each gene on each array (Li, C. & Wong, W.H. Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc. Natl. Acad. Sci. U.S.A. 98, 31-36 (2001)). Keywords = Schwann cells glia Lot batch = 4
