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Sample GSM1211408 Query DataSets for GSM1211408
Status Public on Oct 15, 2013
Title Gata1 24h rep1
Sample type RNA
 
Source name FDCPmix_Gata1 24h
Organism Mus musculus
Characteristics cell line: FDCPmix
infected with: Gata1ERT
Treatment protocol GATA1ERT and Pu.1ERT were subcloned into the pHR-SIN-CSGWEmGFP lentiviral expression construct under control of the SFFV promoter. Gata2 and Pu.1 shRNAs were subcloned into Lentilox 3.7. Recombinant plasmids were packaged into lentiviral particles esssentially by published procedures. FDCPmix cells were transduced at an MOI of between 50 and 200. For gene expression analysis, samples of FDCPmix cells in self-renewal conditions were transduced with lentiviral particles encoding GATA1ERT or PU.1ERT fusion proteins linked to ires-GFP, with empty virus used as a control. GFP+ cells were sorted after 3 days and expanded for a further 7 days under standard conditions, before re-sorting on GFP and addition of 2M 4OH-tamoxifen. Cells were harvested after 0h and 24h of induction and total RNA analyzed by microarray.
Growth protocol FDCPmix were grown in 20% Fischer's medium.
Extracted molecule total RNA
Extraction protocol Total RNA was isolate using TRIzol reagent, and RNA concentration and integrity were determined using RNA 6000 Nano RNA kit (Agilent) on BioAnalyzer 2100 (Agilent).
Label Cy3
Label protocol Total RNA (150ng) and Agilent One-Color RNA Spike-In were reverse-transcribed and linearly amplified in the presence of Cy3-labeled CTP using Low RNA Input One-Color Kit (Agilent) following Agilent protocols, to generate Cy3-labeled cRNA for hybridisation to high-density microarrays. Quality and yield/specific activity of cRNA were determined using RNA 6000 Nano kit and spectrophotometer (NanoDrop ND- 1000). All samples generated comparable quality cRNA profiles, with specific activities in the range of 10.3 to 12.2 pmol Cy3/mg cRNA.
 
Hybridization protocol Each Cy3-labeled cRNA sample (1.6ug as per NanoDrop reading) was hybridised to an individual 44K subarray of 4x44K high-density microarray slide (Whole Mouse Gene Expression Microarrays, Agilent), washed, scanned and feature-extracted as per Agilent protocols.
Scan protocol Scanning and feature extraction was performed as per Agilent protocols
Description ER_10
Data processing The scanned images were analyzed with Feature Extraction Software (Agilent v10.1.1.1) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
 
Submission date Aug 19, 2013
Last update date Oct 15, 2013
Contact name Shamit Soneji
E-mail(s) shamit.soneji@med.lu.se
Organization name BMC
Street address Sölvegatan 19
City Lund
ZIP/Postal code 221 84
Country Sweden
 
Platform ID GPL7202
Series (2)
GSE49990 Dynamic analysis of gene expression and genome wide transcription factor binding during lineage-specification of multipotent progenitors [ER]
GSE49991 Dynamic analysis of gene expression and genome wide transcription factor binding during lineage-specification of multipotent progenitors

Data table header descriptions
ID_REF
VALUE Normalized signal intensity

Data table
ID_REF VALUE
A_52_P756921 3.934
A_52_P1133703 9.308
A_52_P1052870 4.809
A_52_P1021909 6.539
A_51_P133060 3.627
A_51_P287221 6.313
A_52_P112159 4.594
A_51_P378063 4.668
A_52_P289204 6.092
A_51_P439156 6.929
A_52_P2181 7.311
A_52_P53154 1.83
A_52_P527925 7.019
A_52_P315910 5.081
A_52_P519870 7.557
A_52_P96087 6.738
A_51_P137808 7.063
A_52_P552026 8.507
A_52_P42395 5.792
A_52_P350876 5.198

Total number of rows: 41278

Table truncated, full table size 766 Kbytes.




Supplementary file Size Download File type/resource
GSM1211408_US10020348_251486830235_S01_GE1_107_Sep09_1_3.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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