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Transcription profiling of rat Htt14A2.5 (PC12) cells expressing truncated Htt exon 1-EGFP fusion protein


ABSTRACT: 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.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Barbara Apostol 

PROVIDER: E-GEOD-2602 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Mutant huntingtin alters MAPK signaling pathways in PC12 and striatal cells: ERK1/2 protects against mutant huntingtin-associated toxicity.

Apostol Barbara L BL   Illes Katalin K   Pallos Judit J   Bodai Laszlo L   Wu Jun J   Strand Andrew A   Schweitzer Erik S ES   Olson James M JM   Kazantsev Aleksey A   Marsh J Lawrence JL   Thompson Leslie Michels LM  

Human molecular genetics 20051205 2


Huntington's disease (HD) is a devastating neurodegenerative disorder caused by an expanded polyglutamine (polyQ) tract within the huntingtin protein (Htt). Identifying the pathways that are altered in response to the mutant protein is crucial for understanding the cellular processes impacted by the disease as well as for the rational development of effective pharmacological interventions. Here, expression profiling of a cellular HD model identifies genes that implicate altered mitogen-activated  ...[more]

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