Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse mammary C57MG cells transfected with Wnt-1 or empt vector to identify targets of Wnt-1

ABSTRACT: RNA from murine mammary C57MG cells transfected with an empty control vector (PLNCx) or an expression vector encoding Wnt-1 was isolated and analyzed on Affymetrix MG-U74Av2 arrays. Sample preparation was done as recommended by the manufacturer. Images were analyzed using Affymetrix Microarray Suite 4.0 software. Comparison of Wnt-1 expressing and control cells revealed Wnt-1 targets.

ORGANISM(S): Mus musculus

SUBMITTER: Ludger Klein-Hitpass

PROVIDER: E-GEOD-1899 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Novel target genes of the Wnt pathway and statistical insights into Wnt target promoter regulation.

Ziegler Slava S Röhrs Sonja S Tickenbrock Lara L Möröy Tarik T Klein-Hitpass Ludger L Vetter Ingrid R IR Müller Oliver O

The FEBS journal 20050401 7

The Wnt pathway controls biological processes via the regulation of target gene expression. The expression of direct Wnt target genes, e.g. cyclin D1 and MYC, is activated by the transcription factor TCF, which binds to specific sequence motifs in the promoter. Indirect target genes are regulated via transcription regulators, which are targets of the Wnt pathway. As an example, MYC regulates the MYC interacting zinc finger protein-1 (MIZ-1), which is able to inhibit the expression of the indirec ...[more]

PMID: 15794748

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Project description:Bone marrow (BM) cells derived from femurs of C57BL/6 mice were used to generate BM-DCs. For this, BM cells (2x106 cells/10 ml) were seeded on 100 mm2 bacterial dishes (Greiner bio-one, Frickenhausen, Germany). DC culture media (IMDM, 5% [v/v] FCS [PAA, Coelbe, Germany], 100 U/ml penicillin, 100 ug/ml streptomycin [both Gibco, Paisly, UK], 50 uM beta-mercaptoethanol [Roth, Karlsruhe, Germany] 5% [v/v] of murine GM-CSF containing cell culture supernatant was replenished on days 3 and 6 of culture. Aliquots of DC cultures were treated with recombinant human 100 ng/ml WNT-1 (Biovision, Milpitas, CA) for 48h on days 7 or 8 of culture. Each 20 ug of total RNA derived from unstimulated BM-DCs treated for 48 h with WNT-1 (100 ng/ml) or left untreated was used for generation of fluorescecne-labeled cDNA probes (Alexa Fluor 555-aha-dUTP for untreated BM-DCs; Alexa Fluor 647-aha-dUTP for WNT-treated BM-DCs). <br><br><br><br>Prehybridized (5X SSPE,0.1% SDS, and 1% BSA) microarrays (Mouse Whole Genome OneArray Microarray v2; Phalanx Biotech, San Diego, U.S.A.) were hybridized with each 20 pmol of either fluorescence-labeled cDNA probe using OneArray Hybridization Buffer with 18% formamide for 16 h (Lucidea SlidePro Hybridizer; GE Healthcare, Munich, Germany) according to the OneArray hybridization protocol. Washed microarrays were analysed using a ScanArray 4000 system equipped with ScanArray 4.0 software (both Perkin-Elmer; Rodgau-Jugesheim, Germany) at 10 pixel size. Laser intensity (90-100%) and photomultiplier tube sensitivity (60-70%) were varied to optimize signal to background ratios. Derived images weere analyzed using the ScanArray 4.0 easyquant software module. Intensity data were normalized (loess normalization; MIDAS-2.19) and subjected to significance analysis (MEV 4.3) using appropriate modules of the TM4 microarray software suite. Changes in gene expression with p < 5% were used for subsequent analyses. Gene ontology annotations were performed with Panther software (Thomas et al., 2003).

Dataset Information

Transcription profiling of mouse mammary C57MG cells transfected with Wnt-1 or empt vector to identify targets of Wnt-1

Publications

Novel target genes of the Wnt pathway and statistical insights into Wnt target promoter regulation.

Similar Datasets

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