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Sample GSM1000772 Query DataSets for GSM1000772
Status Public on Apr 23, 2013
Title 24h_TSA_rep3
Sample type RNA
 
Channel 1
Source name LD-MEL cells
Organism Mus musculus
Characteristics cell type: Low differentiation-inducible murine erythroleukemia cells, A clone of T-3-Cl-2-0.fl.
treatment: TSA
time: 24h
cell line: T-3-Cl-2-0.fl (Riken Cell No. RCB0561)
Treatment protocol MEL cells were washed in PBS, and stored at -80 °C.
Growth protocol MEL cells were cultured in RPMI1640 medium with 10% fetal calf serum at 37 °C with 5% CO2 concentration.
Extracted molecule total RNA
Extraction protocol Total RNA of MEL cells was extracted using RNeasy Mini Kit (Qiagen).
Label Cy5
Label protocol Twenty mg of the total RNA was labeled with amino-modified dNTP using a SuperScript Indirect cDNA Labeling System (Invitrogen). The amino-modified cDNA was purified with a S.N.A.P column (Invitrogen) and ethanol precipitation. The purified cDNA was labeled with Cye3 Mono-Reactive Dye Pack (Amersham Biosciences) or Cye5 Mono-Reactive Dye Pack (Amersham Biosciences) by coupling reaction. The labeled cDNA was purified with a QIAquick PCR Purification Kit (Qiagen).
 
Channel 2
Source name HD-MEL cells
Organism Mus musculus
Characteristics treatment: TSA
time: 24h
cell type: High differentiation-inducible murine erythroleukemia cells, A clone of T-3-Cl-2-0.fl.
cell line: T-3-Cl-2-0.fl (Riken Cell No. RCB0561)
Treatment protocol MEL cells were washed in PBS, and stored at -80 °C.
Growth protocol MEL cells were cultured in RPMI1640 medium with 10% fetal calf serum at 37 °C with 5% CO2 concentration.
Extracted molecule total RNA
Extraction protocol Total RNA of MEL cells was extracted using RNeasy Mini Kit (Qiagen).
Label Cy3
Label protocol Twenty mg of the total RNA was labeled with amino-modified dNTP using a SuperScript Indirect cDNA Labeling System (Invitrogen). The amino-modified cDNA was purified with a S.N.A.P column (Invitrogen) and ethanol precipitation. The purified cDNA was labeled with Cye3 Mono-Reactive Dye Pack (Amersham Biosciences) or Cye5 Mono-Reactive Dye Pack (Amersham Biosciences) by coupling reaction. The labeled cDNA was purified with a QIAquick PCR Purification Kit (Qiagen).
 
 
Hybridization protocol Microarray was pre-hybrized at 50°C, 60 minutes in 5 x saline sodium citrate buffer (SSC) contained 0.1% sodium dodecyl sulfate (SDS)and 0.1% bovine serum albumin (BSA), washed with water, and rinsed with isopropanol. Mixed Cy3 and Cy5 labeled cDNA solution was denaturation at 95°C, 5 minutes in 5 x SSC contained 1.67 mg/mL acetylated BSA. After cooling, SDS was added to final concentration 0.33%. The microarray was covered with a Gap cover glass (Matsunami), hybridization solution was injected, and the microarray was placed in a hybridization cassette (TeleChem). Hybridization was performed at 50°C for 14 hours. After hybridization, the microarray was washed using a Wash Station (TeleCham) as follow; at 42°C 5 minutes in 2 x SSC contained 0.1%SDS, at room temperature 10 minutes in 0.1 x SSC contained 0.1%SDS, and finally twice in 0.1 x SSC at room temperature for 2min. The microarray was dried by centrifugation.
Scan protocol Hybridization images were scanned using a ScanArray 5000(Perkin-Elmer). Fluorescence intensity of the microarray was quantified using QuantArray 3.0 software (Perkin-Elmer).
Description Technical replicate 3 of 4. 24 hours after 15nM TSA treatment.
control LD-MEL
target HD-MEL
Data processing The microarray data was analyzed using GeneSpring 6.0 (Silicon Genetics). Intensity difference between Cy3 and Cy5 was normalized by LOWESS method.
Signal intensities were higher than negative spot.
 
Submission date Sep 10, 2012
Last update date Apr 24, 2013
Contact name Hitoshi Sasaki
E-mail(s) [email protected]
Organization name Tokyo University of Science
Department Department of Biological Science and Technology
Lab Murakami Laboratory
Street address 2641 Yamazaki
City Noda
State/province Chiba
ZIP/Postal code 278-8510
Country Japan
 
Platform ID GPL16035
Series (1)
GSE40754 Comparison of transcriptome of high differentiation-inducible murine erythroleukemia cells and low differentiation-inducible murine erythroleukemia cells

Data table header descriptions
ID_REF
VALUE Log2 Lowess HD MEL/LD MEL
PRE_VALUE Lowess HD MEL/LD MEL

Data table
ID_REF VALUE PRE_VALUE
H3001A01 -0.2286 0.8534694
H3001A02 0.1522 1.1112672
H3001A03 0.4678 1.3830236
H3001A04 -0.2374 0.84825796
H3001A05
H3001A06 0.0066 1.0045509
H3001A07 0.0561 1.0396751
H3001A08 -0.0316 0.97833335
H3001A09 0.4659 1.3811741
H3001A10 0.1106 1.0797021
H3001A11 0.5650 1.479401
H3001A12 0.3757 1.297473
H3001B01 -1.0589 0.47998932
H3001B02
H3001B03 -0.8013 0.57384914
H3001B04 0.0083 1.0058035
H3001B05 -0.5762 0.6707491
H3001B06 -0.0900 0.9395286
H3001B07 -0.5970 0.6611425
H3001B08 -0.7228 0.6059275

Total number of rows: 22656

Table truncated, full table size 549 Kbytes.




Supplementary file Size Download File type/resource
GSM1000772_TSA_24h-3-part1.txt.gz 110.6 Kb (ftp)(http) TXT
GSM1000772_TSA_24h-3_part2.txt.gz 94.9 Kb (ftp)(http) TXT
Processed data included within Sample table

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