|
| Status |
Public on Apr 25, 2014 |
| Title |
PC3 cells stably overexpressing ERG treated with I-BET762 Replicate 2 PC3_ERG_IBET762_rep2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Total RNA from PC3 cells stably overexpressing ERG treated with I-BET762
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: I-BET762
cell line: PC3
cell type: prostate cancer 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 |
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.
|
| |
|
| Channel 2 |
| Source name |
Total RNA from PC3 cells stably overexpressing ERG treated with DMSO
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: PC3
cell type: prostate cancer 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 |
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.
|
| |
|
| |
| 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.
|
| |
|
| 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 |
|
