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Status |
Public on Oct 31, 2016 |
Title |
LLC4_2cPE |
Sample type |
RNA |
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Source name |
CLL cells - treated with 2cPE, patient LLC4
|
Organism |
Homo sapiens |
Characteristics |
patient id: LLC4 cell type: chronic lymphocytic leukemia (CLL)
|
Treatment protocol |
Leukemia CD19+ B-cells from 10 different patients were treated or not for 6 hours with 6nM of bortezomib or with 4µM of 2cPE
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Growth protocol |
Purified peripheral blood cells were cultured in RPMI-1640 medium supplemented with 10% heated inactivated fetal calf serum. Mediums were supplied with penicillin (100 U/mL), glutamine (2 mmol/L), and streptomycin (100 μg/mL).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated using RNeasy Mini kit (Qiagen). RNA samples concentration and purity (assessed as 260/280 nm and 260/230 nm ratios) were evaluated by NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies; Wilmington, DE), while RNA integrity was assessed by using the Agilent 2100 Bioanalyzer (Agilent Technologies; Waldbrunn, Germany).
|
Label |
biotin
|
Label protocol |
cDNA synthesis and biotin-labeled target synthesis were performed using the GeneAtlas 3´ IVT Express Kit according to the protocol supplied by Affymetrix.
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|
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Hybridization protocol |
Labeled cRNA was hybridized on Affymetrix Human Genome U219 Array.
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Scan protocol |
The HG-U219 Array Strips (Affymetrix; Santa Clara, CA) hybridization, staining, and scanning were performed by using the GeneAtlas Platform.
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Description |
Human Chronic lymphocytic leukemia cells treated with 2cPE
|
Data processing |
Data processing was performed with BioConductor packages (http://www.bioconductor.org/), using Brainarray HGU219_Hs_ENTREZG version 18 custom chip definition file. Robust Multi-Array Average (RMA) normalization was applied.Normalized intensity values for all probes on the Affymetrix chip are presented in the Matrix table. Data analysis was performed using the paired t-test as implemented in the R statistical package. P values were adjusted for multiple testing using the False Discovery Rate method. Differentially expressed genes (DEGs) were selected based on fold change >1.5 and <-1.5 fold and P values <0.05.
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Submission date |
Oct 30, 2015 |
Last update date |
Oct 31, 2016 |
Contact name |
raffaella picco |
E-mail(s) |
raf.picco@uniud.it
|
Organization name |
university of Udine
|
Department |
department of medical and biological sciences
|
Lab |
bioinformatics
|
Street address |
piazzale kolbe 4
|
City |
udine |
ZIP/Postal code |
33100 |
Country |
Italy |
|
|
Platform ID |
GPL20650 |
Series (1) |
GSE74514 |
The isopeptidase inhibitor 2cPE triggers proteotoxic stress and ATM activation in chronic lymphocytic leukemia cells. |
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