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Sample GSM1128385 Query DataSets for GSM1128385
Status Public on Aug 28, 2020
Title HEK RAS transformed UO126 treated rep2
Sample type RNA
 
Source name human embryonic kidney RAS-transformed UO126 treated
Organism Homo sapiens
Characteristics tissue/cell type: embryonic kidney
genotype/variation: RAS transformed
treated with: UO126
Treatment protocol The HRASG12V-transformed human embryonal kidney cells (HA1ER) and their immortalized origin cell line HA1EB were described by [Hahn et al. Nature 400, 464-468 (1999)] . They were cultivated in Minimum Essential Medium Eagle, Alpha Modification (MEM Alpha) from Sigma, supplemented with 10% IFS, 2 mM ultraglutamine, 1% penicillin/streptomycin, 0,1mg/ml hygromycin and 0,5µg/ml puromycin. G418 was freshly added to a final concentration of 0,4 mg/ml . ROSE199, rat ovary epithelial cell line and their KRAS transformed derivative cell line ROSEA25 were described in [Tchernitsa et al. Oncogene 23, 4536-4555 (2004)]. Inducible RAS studies were done in HEK cell line transfected with a constuct bearing Ha-RAS under ER promoter. Cells were treated with tamoxifen before RNA was extracted. U0126 MEK inhibitor was dissolved in DMSO and used in 10µM final concentration for 48h before RNA was extracted.
Growth protocol RAS-ROSE abd ROSE cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) supplemented with 10 % fetal calf serum (FCS) (Sigma-Aldrich), 2 mM Ultraglutamine 1 (Lonza, BioWhittaker), and 100 units/ml penicillin/streptomycin (Biochrom AG) (standard medium). The medium was supplemented with G418 (400 μg/μl). 45000 RAS-ROSE cells were seeded into 6-well plates (BD Falcon) in standard medium 24 h prior to transient transfection of siRNA.
Extracted molecule total RNA
Extraction protocol Total RNA was isolated from cell lines using the mirVanaTM miRNA Isolation Kit from Ambion® (Austin, TX, USA) according to the manufacturer’s instructions. In order to avoid cross-labeling of premature miRNA we used the gel purification option. Subsequently the quality of the small RNA fraction was controlled on a 15% PAAG gel stained with ethidium bromide.
Label biotin
Label protocol Standard Affymetrix labeling protocol
 
Hybridization protocol Standard Affymetrix hybridization protocol
Scan protocol Standard Affymetrix scanning protocol
Description SAMPLE 6
replicate 2
Data processing The data were normalised using RMA normalisation affy package (v.1.32.1) in R/Bioconductor (v.2.14.1).
 
Submission date Apr 23, 2013
Last update date Aug 28, 2020
Contact name Wasco Wruck
E-mail(s) wasco.wruck@med.uni-duesseldorf.de
Organization name Universitätsklinikum Düsseldorf
Street address Moorenstr. 5
City Düsseldorf
ZIP/Postal code 40225
Country Germany
 
Platform ID GPL96
Series (2)
GSE46301 A conserved microRNA signature related to cellular transformation by the RAS oncogene (Affymetrix mRNA)
GSE46806 A conserved microRNA signature related to cellular transformation by the RAS oncogene

Data table header descriptions
ID_REF
VALUE log2 RMA normalized
ABS_CALL

Data table
ID_REF VALUE ABS_CALL
1007_s_at 8.276540332 P
1053_at 8.422762656 P
117_at 5.393706915 A
121_at 9.646417706 P
1255_g_at 3.706500688 A
1294_at 6.68820212 P
1316_at 4.656023119 P
1320_at 5.919901888 P
1405_i_at 5.219144219 P
1431_at 3.886302536 A
1438_at 5.984100878 A
1487_at 6.904392151 M
1494_f_at 6.473250256 M
1598_g_at 9.446916209 P
160020_at 8.452038044 P
1729_at 7.0141951 P
1773_at 5.992899141 A
177_at 5.030775166 M
179_at 8.687685677 A
1861_at 6.736024162 P

Total number of rows: 22283

Table truncated, full table size 540 Kbytes.




Supplementary file Size Download File type/resource
GSM1128385_AS_HEKRAS_UO126_2_08.CEL.gz 2.1 Mb (ftp)(http) CEL
Processed data included within Sample table

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