|
| Status |
Public on Jan 10, 2017 |
| Title |
INA-6 treated with EZH2 inhibitor UNC1999, biological repl 2 |
| Sample type |
RNA |
| |
|
| Source name |
Mature Plasma cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: INA-6
|
| Treatment protocol |
The Multiple Myeloma INA-6 cell line was treated with 1 µM of the EZH2 inhibitior UNC1999 or with 1 µM of the inactive from UNC2400 for 120 hours. Medium and reagents were refershed at 72 hours post-treatment.
|
| Growth protocol |
The Multiple Myeloma INA-6 cell line was cultered in the presence of Intrerleukin-6 (IL-6) in RPMI-1640 AQmediaTM (Sigma) supplemented with 10% FBS (Sigma), 1%GlutaMax™ (Gibco) and antibiotics (penicillin 100U/ml and streptomycin 50 mg/mL; Sigma) at 37°C in a humidified 5% CO2 in-air atmosphere. ). For experimental setup of the 5 days exponentially growing cells were seeded at 100 000 cells/mL overnight before addition of reagents
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIzol® reagent (Invitrogen) and cleaned using the Rneasy MinElute Cleanup Kit (QIAGEN).
|
| Label |
biotin
|
| Label protocol |
The labeling of the RNA for microarray was done according the Affymetrix manufacturer protocol (Affymetrix).
|
| |
|
| Hybridization protocol |
Samples were hybridized using Affymetrix hybridization kit materials. Heat cocktails at 99° for 5 minutes, then 42° for 5 minutes centrifuge at max speed for 1 minute. Transfer 200μl of hyb solution to each array, then tape holes and parafilm. Hybridize 16 hours at 45° at 60rpm,
|
| Scan protocol |
The arrays were scanned using Affymetrix Genechip Scanner.
|
| Description |
MM cell line
|
| Data processing |
The data was processed using GeneSpring v13 software. Where after background correction the data was normalized using quantile normalization and the summarization was done using ExonRMA 16 normilazation.
probe group file: HTA-2_0.r1.pgf
meta-probeset file: HTA-2_0.r1.Psrs.mps
|
| |
|
| Submission date |
Oct 06, 2016 |
| Last update date |
Jan 10, 2017 |
| Contact name |
Prasoon Agarwal |
| E-mail(s) |
helena.jernberg_wiklund@igp.uu.se
|
| Organization name |
Rudbeck Laboratory, Uppsala University
|
| Department |
IGP
|
| Street address |
Dag Hammarshkojlds vag 20
|
| City |
Uppsala |
| State/province |
Sweden |
| ZIP/Postal code |
75185 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL17585 |
| Series (2) |
| GSE87714 |
EZH2 inhibition in multiple myeloma downregulates myeloma associated oncogenes and upregulates microRNAs with potential tumor suppressor functions [mRNA expression] |
| GSE87716 |
EZH2 inhibition in multiple myeloma downregulates myeloma associated oncogenes and upregulates microRNAs with potential tumor suppressor functions |
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