Htt14A2.5 induced verses uninduced

Htt14A2.5 is a clonal rat pheochromocytoma (PC12) cell line, that expresses a truncated form of expanded repeat Htt exon 1 protein fused at the C-terminus to enhanced green fluorescent protein (EGFP). For total RNA preparation, ~3 x 105 cells were plated on 150 mm plates and the following day either induced with 5 microM PA or treated with ethanol alone (uninduced) for 48 hrs followed by direct lysis using the RNeasy total RNA prep as described by the manufacturer (Qiagen, Valencia, CA). Cells were harvested at approximately 40-60% confluency. RNA was quantitated spectrophotometrically and integrity tested by capillary electrophoresis (Agilent 2100 Bioanalyzer). 25 micrograms total RNA was used to generate target cRNAs for hybridization to Affymetrix Rat Genome U34A oligonculeotide arrays (UCI DNA Array Core Facility). Four separate cell growths were performed for the PA induced verses uninduced comparisons. Target cRNA was synthesized separately for each of these cell growths and hybridized individually to the oligonucleotide microarrays. Keywords = huntingtin Keywords = PC12 Keywords: repeat sample