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Sample GSM312106 Query DataSets for GSM312106
Status Public on Sep 28, 2008
Title A9: hr 96 dox (-)
Sample type RNA
 
Source name Ainv18 mouse ES cells engineered to overexpress activated Notch4 upon doxycycline treatment
Organism Mus musculus
Characteristics biological replicate 1
hour = 96
Treatment protocol For reaggregation cultures to support the differentiation of the hematopoietic and vascular lineages, 3 x 10^5 Flk-1+ cells/ml (harvested at d3.5 after addition of serum) were cultured for 24 hours in ultra-low attachment 24-well plates (Corning Costar) with the EB differentiation medium (above) plus 5% Protein-Free Hybridoma Medium-II (PFHM-II, Invitrogen), either in the presence or absence of dox. After 24 hours, the cells were washed and cultured without doxycycline for up to an additional 3 days. Total RNA was harvested from dox-treated or dox-null cultures at 4, 12, 24, 48, or 96 hours.
Growth protocol ES cells were maintained on irradiated feeders in Dulbecco€'s Modified Eagle Medium (DMEM) supplemented with 15% fetal calf serum (FCS), 10% ES cell conditioned medium, penicillin, streptomycin, 1.5 x 10^-4 M monothioglycerol (MTG; Sigma) and LIF (1% conditioned medium). Prior to induction of differentiation, cells were passaged 2 times on gelatin-coated plates in Iscove Modified Dulbecco Medium (IMDM) containing the same supplements mentioned above to deplete the population of feeder cells. ). Prior to induction of differentiation, cells were passaged 2 times on gelatin-coated plates in Iscove Modified Dulbecco Medium (IMDM) containing the same supplements mentioned above to deplete the population of feeder cells. For the generation of EBs, the cells were harvested and cultured in 60 mm low attachment Petri grade dishes (VWR) with IMDM supplemented with 2 mM L-glutamine (Gibco/BRL), 200 µg/mL transferrin (Boehringer Mannheim), 0.5 mM ascorbic acid (Sigma), 4 x 10^-4 M MTG plus 15% FCS.
Extracted molecule total RNA
Extraction protocol Qiagen RNeasy Mini protocol for isolation of total RNA from animal cells, with on column Dnase treatment.
Label Alexa-647
Label protocol see CodeLink Gene Expression System: Manual Labelled cRNA target Preparation
 
Hybridization protocol see CodeLink Gene Expression System: Single-Assay Bioarray Hybridization and Detection
Scan protocol see CodeLink Gene Expression System: Single-Assay Bioarray Hybridization and Detection, using Axon 4000B scanner
Description ***** These records currently present results containing only probes for the Wnt, BMP/TFGb, and Notch pathways as published in Chen et al., 2008, Nature Biotechnology. The full set of processed and raw data tables will be available by June 1, 2009. *****
Data processing median normalization, CodeLink Expression Analysis, filtered for Wnt, BMP/TFGb, and Notch pathway probesets
 
Submission date Aug 12, 2008
Last update date May 29, 2009
Contact name Robert A Stull
E-mail(s) bobstullca@gmail.com
Phone (650)244-9990
Organization name VistaGen Therapeutics
Department Genomics
Street address 384 Oyster Point Blvd, Suite #8
City South San Francisco
State/province CA
ZIP/Postal code 94080
Country USA
 
Platform ID GPL2897
Series (1)
GSE12425 Dox-regulated Notch4 overexpression in mouse ES cells redirects hemagioblasts to a cardiac fate.

Data table header descriptions
ID_REF
VALUE median normalized signal intensity value

Data table
ID_REF VALUE
1001 131.4813
1002 17.0934
1003 0.229
1004 1.0147
1005 0.7162
1006 0.2133
1007 134.1221
1008 133.114
1010 2.1745
1012 1.0942
1013 0.5879
1014 135.6554
1015 135.56
1017 0.2986
1018 0.0775
1019 0.109
1020 0.8207
1021 137.1846
1022 133.0654
1024 0.2252

Total number of rows: 36227

Table truncated, full table size 488 Kbytes.




Supplementary file Size Download File type/resource
GSM312106.TXT.gz 1.4 Mb (ftp)(http) TXT
Processed data included within Sample table

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