Expression analysis of neurospheres generated in vitro

Neurospheres generated in vitro were treated with non-epinephrine or potassium chloride. Gene expression analysis was then carried out to identify genes that are up or down regulated due to chemical treatement. The microarray data for the neurosphere samples was generated at Queensland Brain Institute. Briefly, RNA was isolated from neurospheres generated in presence of either N (norepinephrine) or K (potassium chloride)and converted to cDNA using Applause WT-Amp ST kit (NuGEN). Fragmentation and biotin labelling of the cDNA was carried out using Encore Biotin module (NuGEN) for Affymetrix GeneChip Mouse Gene 1.0 ST arrays. Labelled cDNA was hybridised to arrays in an oven at 45°C. Hybridised arrays were scanned using the Affymetrix GeneChip Scanner and the scanned data (.CEL files) was transferred to the Partek Genomics Suite and data analysis was carried out as described in Narayanan RK, Mangelsdorf M, Panwar A, Butler TJ, Noakes PG, et al. (2013).