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Status |
Public on Jul 26, 2018 |
Title |
M0 CM treated A549 |
Sample type |
RNA |
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Source name |
M0 macrophage conditioned media
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Organism |
Homo sapiens |
Characteristics |
cell line: A549 cell type: lung cancer
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Treatment protocol |
A549 cells were treated with M0, M1, and M2 conditioned media.
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Growth protocol |
cultivated in RPMI + 5% fetal bovine serum
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Extracted molecule |
total RNA |
Extraction protocol |
Total RNA were extracted by RNeasy mini kit (Cat. 74106; Qiagen, Hilden, Germany), and 100 ng to 1 ug of total RNA were subjected into reverse transcription using the SuperScriptTM III Reverse transcriptase kit (Invitrogen) as directed by the manufacturer.
|
Label |
CY3-CTP
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Label protocol |
0.2 μg of total RNA was amplified by a Low Input Quick-Amp Labeling kit (Agilent Technologies, USA) and labeled with Cy3 (CyDye, Agilent Technologies, USA) during the in vitro transcription process. 0.6 μg of Cy3-labled cRNA was fragmented to an average size of about 50-100 nucleotides by incubation with fragmentation buffer at 60°C for 30 minutes.
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Hybridization protocol |
Correspondingly fragmented labeled cRNA is then pooled and hybridized to Agilent SurePrint G3 Human V2 GE 8×60K Microarray (Agilent Technologies, USA) at 65°C for 17 h.
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Scan protocol |
After washing and drying by nitrogen gun blowing, microarrays are scanned with an Agilent microarray scanner (Agilent Technologies, USA) at 535 nm for Cy3
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Description |
2 days treatment
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Data processing |
Scanned images are analyzed by Feature extraction10.5.1.1 software (Agilent Technologies, USA), an image analysis and normalization software is used to quantify signal and background intensity for each feature.
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Submission date |
Mar 23, 2017 |
Last update date |
Jul 26, 2018 |
Contact name |
Chih-Chan Lee |
E-mail(s) |
chihchanlee@gmail.com
|
Phone |
+886-952-520047
|
Organization name |
Molecular medicine program
|
Street address |
No.155, Sec.2, Linong Street
|
City |
Taipei |
ZIP/Postal code |
112 |
Country |
Taiwan |
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|
Platform ID |
GPL17077 |
Series (1) |
GSE96943 |
Expression analysis of A549 cells following treatment with conditioned medium from polarized macrophages |
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