Unknown,Transcriptomics,Genomics,Proteomics

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Expression analysis of uninfected Boechera stricta and infected Boechera stricta with Puccinia monoica forming pseudoflowers using an Arabidopsis thaliana gene expression 4x72K array design from NimbleGen.

ABSTRACT: The expression of 30362 plant genes from uninfected flowers of Boechera stricta, uninfected steam and leaves of B. stricta and infected B. stricta with Puccinia monoica forming pseudoflowers. We hybridized cDNA from each sample to an Arabidopsis thaliana gene expression 4x72K format NimbleGen array (ATH6_60mer_expr). We used a eukaryotic gene expression array design No.5048 from NimbleGen (Cat No. A4511001-00-01). Each 5048 array measures the expression level of 30,362 target genes from Arabidopsis thaliana in a 4-plex format 4x72K with with 72,000 probes per array, a total of two probes per target gene, and 60-mer probe length. Total RNA samples recovered from infected leaves of Boechera stricta with Puccinia monoica (pseudoflowers) and uninfected stem and leaves of B. stricta. Experiments included three/two biological repllicates from each sample. We carried out total RNA extractions for all samples using RNAesy Plant Mini Kit (Qiagen, Cat No. 74904). cDNA synthesis was performed by NimbleGen.

ORGANISM(S): Boechera stricta

SUBMITTER: Liliana Cano

PROVIDER: E-GEOD-41165 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Glioblastoma multiforme (GBM) is the most prevalent type of adult brain tumor, and one of the deadliest tumors known to mankind. The genetic understanding of GBM is, however, limited, and the molecular mechanisms which facilitate GBM cell survival and growth within the tumor microenvironment are largely unknown. We applied digital karyotyping and single nucleotide polymorphism (SNP) arrays to screen for copy number changes in GBM samples and found that the most frequently amplified region is at chromosome 7p11.2. The high resolution of digital karyotyping and SNP arrays permits the precise delineation of amplicon boundaries and has enabled identification of the minimal region of amplification at 7p11.2, which contains two genes, EGFR and SEC61?. SEC61? encodes a subunit of a heterotrimeric protein channel located in the endoplasmic reticulum (ER). In addition to its high frequency of gene amplification in GBMs, SEC61? is also remarkably overexpressed in 77% of GBMs, but not in lower-grade gliomas. The siRNA-mediated knockdown of SEC61? expression in tumor cells led to growth suppression and apoptosis. Furthermore, we showed that pharmacological ER stress agents induce SEC61? expression in GBM cells. Together, these results indicate that aberrant expression of SEC61? serves significant roles in GBM cell survival, likely via a mechanism that is involved in the cytoprotective ER stress-adaptive response to the tumor microenvironment. hSETD1A was silenced in HCT116 cells using retrovirus harboring shRNA specifically against the hSETD1A gene. 10?g RNA was reverse transcribed to cDNA using the Applied Biosystems High Capacity cDNA Reverse Transcription kit according to the manufacturer’s instructions (Applied Biosystems). cDNA products were treated with 100ng RNaseA and then purified using the Qiagen PCR purification kit according to the manufacturer’s instructions. NimbleGen Human Gene Expression array was purchased from Roche Applied Sciences. cDNA samples were labeled, Cy3 hybridized, and processed at the FSU NimbleGen Microarray Facility at Florida State University.

Dataset Information

Expression analysis of uninfected Boechera stricta and infected Boechera stricta with Puccinia monoica forming pseudoflowers using an Arabidopsis thaliana gene expression 4x72K array design from NimbleGen.

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