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HEK293 stably expressing Autoimmune regulator protein

ABSTRACT: Auto Immune REgulator (AIRE) protein expression in HEK293 cells Three cell-lines where used in the study: HEK293-NC; control cell-line expressing yellow fluorescent protein (YFP) HEK-AIRE1, HEK-AIRE2; two different clone-derived Autoimmune regulator protein expressing cell-lines HEK-AIRE-D312A; cell line expressing Autoimmune regulator protein with D312A mutation in the first PHD finger With all cell-lines three independent experiments were performed.

ORGANISM(S): Homo sapiens

SUBMITTER: PÃ¤rt Peterson

PROVIDER: E-GEOD-16877 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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AIRE activated tissue specific genes have histone modifications associated with inactive chromatin.

Org Tõnis T Rebane Ana A Kisand Kai K Laan Martti M Haljasorg Uku U Andreson Reidar R Peterson Pärt P

Human molecular genetics 20090910 24

The Autoimmune Regulator (AIRE) protein is expressed in thymic medullary epithelial cells, where it promotes the ectopic expression of tissue-restricted antigens needed for efficient negative selection of developing thymocytes. Mutations in AIRE cause APECED syndrome, which is characterized by a breakdown of self-tolerance. The molecular mechanism by which AIRE increases the expression of a variety of different genes remains unknown. Here, we studied AIRE-regulated genes using whole genome expre ...[more]

PMID: 19744957

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Homo sapiens

Project description:HEK293 stably expressing Autoimmune regulator protein

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ChIP to AIRE regulated genes in HEK 293 cells

Project description:The Autoimmune Regulator (AIRE) protein is expressed in thymic medullary epithelial cells, where it promotes the ectopic expression of tissue-restricted antigens needed for efficient negative selection of developing thymocytes. Mutations in AIRE cause APECED syndrome, which is characterized by a breakdown of self-tolerance. The molecular mechanism by which AIRE increases the expression of a variety of different genes remains unknown. Here, we studied AIRE-regulated genes using whole genome expression analysis and chromatin immunoprecipitation. We show that AIRE preferentially activates genes that are tissue-specific and characterized by low levels of initial expression in stably transfected HEK293 cell model and mouse thymic medullary epithelial cells. In addition, the AIRE-regulated genes lack active chromatin marks, such as histone H3 trimethylation (H3K4me3) and acetylation (AcH3), on their promoters. We also show that during activation by AIRE, the target genes acquire histone H3 modifications associated with transcription and RNA polymerase II. In conclusion, our data show that AIRE is able to promote ectopic gene expression from chromatin associated with histone modifications characteristic to inactive genes. We compared the gene expression profiles of HEK293 cell-lines expressing either YFP (NC) or AIRE (AIRE1) proteins using Illumina BeadChips (GSE16877). Based on these results we chose more than 50 genes that where upregulated in the presence of AIRE and used chromatin immunoprecipitation and custom made Nimblegen tiling arrays to study the regions of 200kb up-and downstream of the genes in HEK293 cells expressing either YFP or AIRE protein. The following antibodies where used in the ChIP experiments: H3K4me3, AcH3, H3K27me3, H3, PolII and AIRE.

2010-09-10 | E-GEOD-17216 | biostudies-arrayexpress

HEK293 stably expressing Autoimmune regulator protein

Project description:Auto Immune REgulator (AIRE) protein expression in HEK293 cells

2009-09-11 | GSE16877 | GEO

Project title: Endogenous AIRE-immunoprecipitation from primary human neutrophils

Project description:Autoimmune regulator (AIRE) is a transcriptional regulator. We here assess the role for AIRE in primary human neutrophils. We found that the functional role for AIRE in neutrophils is to regulate the extrinsic apoptotic pathway involving the Fas/TNFR death receptors and Cathepsin G and the expression of unique immune mediators.

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Gene expression of HEK 293 cells transfected with Vector (PCMV), Aire, or MBD-VP16

Project description:HEK 293 cells were transiently transfected with plasmids expressing Vector only(PCMV), Aire, or MBD-VP16 with the goal of comparing the global gene expression profiles in the Aire and MBD-VP16 groups We used microarrays to detail the global gene expression profile. HEK 293 cells were transiently transfected with plasmids expressing Vector only(PCMV), Aire, or MBD-VP16 and 72 hrs post transfection total RNA was collected for microarray analysis.

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ChIP to AIRE regulated genes in HEK 293 cells

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The effect of the autoimmune regulator (Aire) gene on the transcriptional expression profile of 3.10 mTEC cell line.

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