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Sample GSM1967610 Query DataSets for GSM1967610
Status Public on Dec 09, 2015
Title FCCP Replicate 2
Sample type RNA
 
Channel 1
Source name C3A cells exposed 2h
Organism Homo sapiens
Characteristics cell line: C3A
Treatment protocol C3A cells were exposed for 2 hours to substances. For microarray analysis, 6 replicate wells per concentration were pooled. DMSO (0.1%) was used as solvent control.
Growth protocol C3A cells were grown in 75 cm2 culture flasks until approx. 80% confluency. Cells were then subcultured as quoted above until the uptake of trypsinised cells in media. C3A cells were diluted to reach a final density of 250 000 cells/ml. 200 µl of cell suspension were seeded in the inner 60 wells of a black 96-well plate with clear bottom (Greiner bio-one, Product 655906, Reinach, Switzerland). The outer rows were filled with 200 µl 1x PBS (phosphate buffered saline, Sigma-Aldrich, Buchs, Switzerland). Cells were pre-incubated at 37°C for 24 h. After pre-incubation cells were dosed with the test chemicals.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted using RNeasy Mini Kit (QUIAGEN, Product 74104, Hombrechtikon, Switzerland) according to manufacturers’ instructions. DNA was digested using the RNeasy Mini Kit optional on-column DNase digestion. Quality of extracted RNA was checked via the Nanodrop 1000 spectraphotomoeter V3.7, as well as running the samples on a 1.5% agarose gel.
Label cy5
Label protocol The RNA 6000 NanoChip kit (Agilent Technologies, number 506791511) was used with the Agilent 2100 Bioanalyzer for analysis of total RNA samples following the manual instructions. Samples with a RIN (RNA integrity number) above 7 were considered appropriate for consequent microarray testing. For Microarray analysis Agilent SurePrint G3 Human Gene Expression Microarrays (8x60K) were used. All replicates of the same treatment were labeled with the same dye (either Cy3 or Cy5). Each dye (Cy3/Cy5) had its own DMSO control.
 
Channel 2
Source name C3A cells control
Organism Homo sapiens
Characteristics cell line: C3A
Treatment protocol C3A cells were exposed for 2 hours to substances. For microarray analysis, 6 replicate wells per concentration were pooled. DMSO (0.1%) was used as solvent control.
Growth protocol C3A cells were grown in 75 cm2 culture flasks until approx. 80% confluency. Cells were then subcultured as quoted above until the uptake of trypsinised cells in media. C3A cells were diluted to reach a final density of 250 000 cells/ml. 200 µl of cell suspension were seeded in the inner 60 wells of a black 96-well plate with clear bottom (Greiner bio-one, Product 655906, Reinach, Switzerland). The outer rows were filled with 200 µl 1x PBS (phosphate buffered saline, Sigma-Aldrich, Buchs, Switzerland). Cells were pre-incubated at 37°C for 24 h. After pre-incubation cells were dosed with the test chemicals.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted using RNeasy Mini Kit (QUIAGEN, Product 74104, Hombrechtikon, Switzerland) according to manufacturers’ instructions. DNA was digested using the RNeasy Mini Kit optional on-column DNase digestion. Quality of extracted RNA was checked via the Nanodrop 1000 spectraphotomoeter V3.7, as well as running the samples on a 1.5% agarose gel.
Label cy3
Label protocol The RNA 6000 NanoChip kit (Agilent Technologies, number 506791511) was used with the Agilent 2100 Bioanalyzer for analysis of total RNA samples following the manual instructions. Samples with a RIN (RNA integrity number) above 7 were considered appropriate for consequent microarray testing. For Microarray analysis Agilent SurePrint G3 Human Gene Expression Microarrays (8x60K) were used. All replicates of the same treatment were labeled with the same dye (either Cy3 or Cy5). Each dye (Cy3/Cy5) had its own DMSO control.
 
 
Hybridization protocol After hybridization, the microarrays were washed using Gene Expression Wash Buffer Kit (Agilent Technologies),
Scan protocol Arrays were scanned using the Scan Control Software A8.5 with the feature extraction 10.X Software (Agilent technologies) on the DNA microarray scanner G2505C (Agilent Technologies).
Data processing M values (log2FoldChange) were calculated and LOEWESS and centering normalization applied. Non uniform and control spots were removed from the data set before duplicated spots on the array were averaged (Median). Then exposure replicates were averaged using median. To identify Significantly regulated genes genes expressed |M|>2 were selected and at-test performed. Adjusted p-values were calculated using the Bonferroni-Hochberg correction and a cut-off of 0.1 applied.
 
Submission date Dec 08, 2015
Last update date Dec 09, 2015
Contact name Jessica Legradi
E-mail(s) jessica.legradi@vu.nl
Organization name VU Amsterdam
Department IVM
Street address De Boelelaan 1087
City Amsterdam
ZIP/Postal code 1081 HV
Country Netherlands
 
Platform ID GPL13607
Series (2)
GSE75782 C3A Cells: Exposure vs Control [two-channel arrays]
GSE75784 Mode of toxic action assignment of chemicals

Data table header descriptions
ID_REF
VALUE normalized log2 ratio representing exposure/control

Data table
ID_REF VALUE
1 0
2 0
3 0
4 0.379853837
5 0.421846623
6 -1.105721229
7 -0.542153603
8 0.131326188
9 0.022946956
10 0.524973824
11 0.712900358
12 -0.967470621
13 -0.787190991
14 -0.853423904
15 -0.05072697
16 0.207471082
17 0.488839356
18 -0.508599528
19 -0.274818123
20 0.977472066

Total number of rows: 62976

Table truncated, full table size 911 Kbytes.




Supplementary file Size Download File type/resource
GSM1967610_US22502676_252800418273_S01_GE2_107_Sep09_2_3.txt.gz 21.0 Mb (ftp)(http) TXT
Processed data included within Sample table

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