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Status |
Public on Apr 25, 2014 |
Title |
RWPE cells stably overexpressing ERG treated with JQ1 Replicate 2 RWPE_ERG_JQ1_rep2_swap |
Sample type |
RNA |
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|
Channel 1 |
Source name |
Total RNA from RWPE cells stably overexpressing ERG treated with JQ1
|
Organism |
Homo sapiens |
Characteristics |
treatment: JQ1 cell line: RWPE cell type: Benign prostate cells transduced with: ERG lentivirus genotype/variation: AR negative
|
Growth protocol |
Cells were cultured in ATCC recommended media supplemented with 10% FBS.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using miRNeasy Kit (Invitrogen) following to the manufacturer’s instructions.
|
Label |
Cy3
|
Label protocol |
1 µg of total RNA were primed with 2 µl of 100 µM T7-oligo-dT DNA primer at 65°C for 10 min, then reversed transcribed at 40°C for 2 h in the presence of 400 U MMLV RTase (Agilent), and labeled with T7 polymerase with 100 µM each of dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy5-label or Cy3-label.
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Channel 2 |
Source name |
Total RNA from RWPE cells stably overexpressing ERG treated with DMSO
|
Organism |
Homo sapiens |
Characteristics |
cell line: RWPE cell type: Benign prostate cells transduced with: ERG lentivirus
|
Growth protocol |
Cells were cultured in ATCC recommended media supplemented with 10% FBS.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using miRNeasy Kit (Invitrogen) following to the manufacturer’s instructions.
|
Label |
Cy5
|
Label protocol |
1 µg of total RNA were primed with 2 µl of 100 µM T7-oligo-dT DNA primer at 65°C for 10 min, then reversed transcribed at 40°C for 2 h in the presence of 400 U MMLV RTase (Agilent), and labeled with T7 polymerase with 100 µM each of dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy5-label or Cy3-label.
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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 |
Scanned on an Agilent G2505B scanner. Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1).
|
Data processing |
Agilent Feature Extraction Software (v 8.5.1.1) was used for background subtraction and LOWESS normalization.
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Submission date |
Feb 14, 2014 |
Last update date |
Apr 26, 2014 |
Contact name |
Marcin Piotr Cieslik |
E-mail(s) |
marcin.cieslik@gmail.com
|
Organization name |
University of Michigan
|
Department |
Pathology
|
Lab |
MCTP
|
Street address |
500 S State St
|
City |
Ann Arbor |
State/province |
Michigan |
ZIP/Postal code |
48104 |
Country |
USA |
|
|
Platform ID |
GPL4133 |
Series (2) |
GSE55063 |
Therapeutic Targeting of BET Bromodomain Proteins in Castration-Resistant Prostate Cancer (microarray) |
GSE55064 |
Therapeutic Targeting of BET Bromodomain Proteins in Castration-Resistant Prostate Cancer |
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