Systemic and intrinsic functions of ATRX in glial cell commitment and CNS myelination

Total forebrain RNA (10 μg) was isolated from three P17 pairs of littermate-matched AtrxFoxG1Cre and control mice using the RNeasy Mini kit (Qiagen Cat# 74104). cRNA was generated and hybridized to an Affymetrix Mouse Genome 430 2.0 Array at the London Regional Genomics Center (London, Canada). Probe signal intensities were generated using GCOS1.4 (Affymetrix Inc., Santa Clara, CA) using default values for the Statistical Expression algorithm parameters and a Target Signal of 150 for all probe sets and a Normalization Value of 1. Gene level data was generated using the RNA preprocessor in GeneSpring GX 7.3.1 (Agilent Technologies Inc., Palo Alto, CA). Data were then transformed (measurements less than 0.01 set to 0.01), normalized per chip to the 50th percentile, and per gene to control samples. Probe sets representing Atrx transcripts were removed (10 sets). The remaining probe sets were filtered by fold change ≥1.5 between control and AtrxFoxG1Cre samples, and by confidence level of P<0.05. Significantly overrepresented GO categories were determined using the Enrichr web platform 3 controls and 3 ATRX FOXG1 Cre Knockouts