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Gene expression data from cultured mouse cortical (mCTX) neurons in different stimulation and knockdown conditions

ABSTRACT: Although the induction of C-FOS in the brain has been extensively studied for several decades to date there has been no attempt to identify the targets of C-FOS at a genome wide level, and it was not known how many genes C-FOS activates in a given cell. To identify potential C-FOS target genes, we performed microarray analysis on RNA obtained from mouse cortical (mCTX) neurons infected with lentivirus containing either a control shRNA (targeting firefly luciferase) or c-Fos shRNA that were subsequently depolarized with 0, 1, 3, or 6 hours of KCl. Embryonic day 16.5 cortices from C57BL/6 mice were dissected, dissociated, plated, and subsequently grown in a cell culture incubator that maintained a temperature of 37 degrees C and a CO2 concentration of 5%. On DIV5, lentiviral supernatant containing either a lentivirus expressing a control shRNA targeting firefly luciferase or a lentivirus expressing shRNA targeting c-Fos was applied to cultures for 6hrs with 6ug/mL polybrene. At DIV7, mCTX neurons were silenced and stimulated with KCl for either 0, 1, 3, or 6hrs, and total RNA was purified using Trizol and the RNeasy mini kit from Qiagen. 10ug of labeled cDNA was subsequently hybridized to Affymetrix mouse genome 430 2.0 arrays with affymetrix processing.

ORGANISM(S): Mus musculus

SUBMITTER: Athar Malik

PROVIDER: E-GEOD-60049 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene expression data from cultured mouse cortical (mCTX) neurons in different stimulation and knockdown conditions

Project description:Although the induction of C-FOS in the brain has been extensively studied for several decades to date there has been no attempt to identify the targets of C-FOS at a genome wide level, and it was not known how many genes C-FOS activates in a given cell. To identify potential C-FOS target genes, we performed microarray analysis on RNA obtained from mouse cortical (mCTX) neurons infected with lentivirus containing either a control shRNA (targeting firefly luciferase) or c-Fos shRNA that were subsequently depolarized with 0, 1, 3, or 6 hours of KCl.

2014-09-07 | GSE60049 | GEO

Transcription profiling of mouse naive CD4+CD25-CD62Lhi T cells and nTreg cells with Eos knock-down

Project description:Eos expression in Treg cells have been documented by several microarrays including ours. We hypothesized that Eos facilitates Foxp3- dependent gene repression in Regulatory T cells. In order to investigate the role of Eos in mediating the Foxp3-dependent gene silencing program, we utilized lentiviral shRNA knockdown of Eos in natural Tregs isolated from the periphery of Balb/C mice. A renilla luciferase (RL) gene specific shRNA lentivirus was used as a control for the transduction of cells. The transcriptional profile of naive T cells, natural Tregs, Eos knockdown Tregs, and control shRNA knockdown Tregs was compared using Agilent 4x 44K whole mouse genome array. The goal of this microarray is to document the global effect of the loss of Eos expression on the transcriptional profile of natrual Treg cells. Experiment Overall Design: Eos knock-down (si-Eos) was mediated by Lentivirus expressing GFP as a reporter of transduction and sorting marker. Renilla luciferase specific shRNA lentiviral transduction was carried out in parallel as a control. NaÃ¯ve (CD4+CD25-CD62Lhi) T cells and nTreg cells were freshly isolated from Balb/C mice.

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YAP conveys an invasive phenotype to clear cell renal cell carcinoma cell lines via endothelin-2

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PTHR1 knock-down in osteosarcoma cells

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