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Sample GSM1108292 Query DataSets for GSM1108292
Status Public on May 21, 2018
Title HMVEC-L_replicative senescent_rep2
Sample type RNA
Source name HUVEC_PD > 20_replicate2
Organism Homo sapiens
Characteristics cell type: replicative senescent HMVEC-L
tissue: pulmonary microvascular
Treatment protocol TNF-α (Sigma-Aldrich, USA) was used to induce premature senescence in HMVEC-L. The cells were subjected to fifteen successive sub-lethal treatments of 20 ng/ml TNF-α in EGM-2MV containing 5% FBS, with one treatment per day for fifteen consecutive days. Control cultures were maintained in parallel without TNF-α.
Growth protocol HMVEC-L was purchased from Lonza (Walkersville, MD, USA) and maintained in Microvascular Endothelial Cell Growth Medium-2 (EGM-2MV) (Lonza), at 37oC in a humidified atmosphere of 95% air/5% CO2. Young and RS cells used in experiments are cells with PD < 5 and PD > 20 respectively.
Extracted molecule total RNA
Extraction protocol Total RNAs were isolated from cells using miRNeasy mini kit (Qiagen) according to manufacturer’s protocol. RNA was quantified using Nanophotometer (Implen GmbH, Germany). RNA integrity was accessed using Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA, USA).
Label Cy3
Label protocol Total RNAs (100ng) were labeled with Cyanine 3-pCp using miRNA Complete Labeling and Hyb Kit (Agilent Technologies Inc., USA) according to manufacturer’s protocol. After labeling, Cy3-labelled RNAs were dried completely using vacuum concentrator (Genevac, USA) at 45oC to 55oC for 2 to 3 hours.
Hybridization protocol 18 µl nuclease free water was added to resuspend the dried Cy3-labelled RNAs. 4.5 µl of the 10x GE Blocking Agent and 22.5 µl of 2x Hi-RPM Hybridization Buffer were added to each sample and incubated 100°C for 5 minutes. 45 µl mixture was then hybridized to Agilent Human MicroRNAs Miroarrays (G4770C) for 20 hours at at 55°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 5 minutes with moderate stirring at room temperature with GE Wash Buffer 1 (Agilent) and 5 minute with moderate stirring at 37°C GE Wash buffer 2 (Agilent).
Scan protocol Slides were scanned immediately after washing on the Agilent Microarray Scanner C using one color scan setting for the 8x15K array slides (Scan Area 61x21.6 mm, Scan resolution 5μm, Dye channel is set to Green and output is 20 bit TIFF).
Description miRNA expression of replicative senescent HMVEC-L
Data processing The scanned images were analyzed with Feature Extraction Software (Agilent) using protocol: miRNA_107_Sep09 and Grid: 021827_D_F_20110707.
Submission date Mar 27, 2013
Last update date May 21, 2018
Contact name Pooi-Fong Wong
Organization name University of malaya
Department Pharmacology
Street address Faculty of Medicine
ZIP/Postal code 50603
Country Malaysia
Platform ID GPL14767
Series (2)
GSE45539 miRNA signature of young, replicative and TNF-α-induced premature senescent of human microvascular endothelial cells-lung (HMVEC-L)
GSE45541 Young, replicative and TNF-a-induced premature senescent of human microvascular endothelial cells-lung (HMVEC-L)

Data table header descriptions
VALUE Agilent Feature Extraction default gProcessedSignal

Data table
1 1.56E+04
2 1.54E+01
3 4.04E+03
4 -4.51E-01
5 1.03E+01
6 -1.40E-01
7 -1.23E+00
8 1.94E-01
9 9.30E+00
10 2.41E+00
11 3.99E+00
12 3.91E+00
13 4.27E+00
14 1.97E+00
15 1.97E+00
16 2.72E+00
17 2.20E+00
18 9.46E-01
19 2.26E+00
20 2.53E+00

Total number of rows: 15739

Table truncated, full table size 228 Kbytes.

Supplementary file Size Download File type/resource
GSM1108292_US91203660_252182712938_S01_miRNA_107_Sep09_1_2.txt.gz 839.6 Kb (ftp)(http) TXT
Processed data included within Sample table
Processed data provided as supplementary file

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