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Sample GSM1230233 Query DataSets for GSM1230233
Status Public on Sep 13, 2013
Title Aiolos-overexpressing A549 cells Replicate 1
Sample type RNA
 
Channel 1
Source name control vector-expressing A549 cells
Organism Homo sapiens
Characteristics cell line: A549
cell type: Human lung adenocarcinoma epithelial cells
Growth protocol A549 cells were grown in RPMI1640 medium with 10% FBS, transfected with control vector or Aiolos expression vector.
Extracted molecule total RNA
Extraction protocol Total RNA extracted using Trizol following manufacturer's instructions
Label Cy3
Label protocol 100ng of total RNA was used to produce polyadenylated RNA with polyA polymerase (Ambion, Austin, TX, USA).The products were transcribed to single-stranded DNA witholigo-dT primers. RNA was selectively degraded by RNase H, leaving RNA oligomers as primers for a DNA polymerase to synthesize double-stranded DNA(dsDNA). The dsDNA was purified and served as a template to generate antisense cDNA by cRNA reverse transcription.The fluorescence labeling reactions contained 5× Klenow buffer, dNTP (120 mM dATP, 120 mM dGTP, 120 mM dTTP, and 60 mM dCTP), 40 mM Cy5-dCTP or Cy3-dCTP (GE Healthcare) and Klenow enzyme (TaKaRa, Dalian, China). The reaction was carried out at 37°C for 90 min and stopped by addition of 25 mM EDTA and 0.25% sodium dodecyl sulfate (SDS).
 
Channel 2
Source name Aiolos-overexpressing A549 cells
Organism Homo sapiens
Characteristics cell line: A549
cell type: Human lung adenocarcinoma epithelial cells
Growth protocol A549 cells were grown in RPMI1640 medium with 10% FBS, transfected with control vector or Aiolos expression vector.
Extracted molecule total RNA
Extraction protocol Total RNA extracted using Trizol following manufacturer's instructions
Label Cy5
Label protocol 100ng of total RNA was used to produce polyadenylated RNA with polyA polymerase (Ambion, Austin, TX, USA).The products were transcribed to single-stranded DNA witholigo-dT primers. RNA was selectively degraded by RNase H, leaving RNA oligomers as primers for a DNA polymerase to synthesize double-stranded DNA(dsDNA). The dsDNA was purified and served as a template to generate antisense cDNA by cRNA reverse transcription.The fluorescence labeling reactions contained 5× Klenow buffer, dNTP (120 mM dATP, 120 mM dGTP, 120 mM dTTP, and 60 mM dCTP), 40 mM Cy5-dCTP or Cy3-dCTP (GE Healthcare) and Klenow enzyme (TaKaRa, Dalian, China). The reaction was carried out at 37°C for 90 min and stopped by addition of 25 mM EDTA and 0.25% sodium dodecyl sulfate (SDS).
 
 
Hybridization protocol Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential.
Scan protocol Slides were scanned with an Agilent scanner Type C.
Description Biological replicate 1 of 3. Gene expression changes in Aiolos-overexpressing A549 cells versus control vector-expressing A549 cells
Data processing Images were quantified using Agilent Feature Extraction Software (version 11).
 
Submission date Sep 12, 2013
Last update date Sep 13, 2013
Contact name Zhenyi Ma
E-mail(s) zhyma@tmu.edu.cn
Phone 86-22-83336533
Organization name Tianjin Medical University
Department Biochemistry and Molecular Biology
Street address 22 Qixiangtai Road
City Tianjin
ZIP/Postal code 300070
Country China
 
Platform ID GPL17077
Series (1)
GSE50812 Aiolos promotes anchorage independence by silencing p66Shc transcription in lung carcinoma A549 cells

Data table header descriptions
ID_REF
VALUE normalized log2 ratio (Cy5/Cy3) representing test/reference.

Data table
ID_REF VALUE
A_33_P3901921 -1.0890551
A_33_P3897734 -0.11933422
A_33_P3897698 0.14892864
A_33_P3893191 -0.76745033
A_33_P3892710 -0.84132576
A_33_P3892608
A_33_P3891747 -0.044487953
A_33_P3889862 -0.21012688
A_33_P3889634 -0.6816597
A_33_P3889179 0.12756014
A_33_P3888629 1.0170274
A_33_P3888568 0.22187757
A_33_P3888485 2.9183283
A_33_P3888365 0.6224823
A_33_P3887888 -0.90560055
A_33_P3887081 1.0829291
A_33_P3886938
A_33_P3886737 -0.25869036
A_33_P3886707 0.8158989
A_33_P3885807 -0.19675112

Total number of rows: 50599

Table truncated, full table size 1100 Kbytes.




Supplementary file Size Download File type/resource
GSM1230233_A549Aiolos-rep1.txt.gz 21.7 Mb (ftp)(http) TXT
Processed data included within Sample table

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