|
| Status |
Public on May 08, 2018 |
| Title |
Macrophage GM-CSF + siControl replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Macrophage GM-CSF + siControl
|
| Organism |
Homo sapiens |
| Characteristics |
donor: 1
cell type: GM-CSF Macrophage
genotype/variation: control
|
| Treatment protocol |
Cells were were treated with GM-CSF (1000 U/ml) every 48 hours
|
| Growth protocol |
Freshly isolated CD14+ monocytes were cultured for 7 days in RPMI 10% FCS containing GM-CSF and, after 7 days were transfected with siRNA for PPARG (siPPARG) or siRNA with a scramble sequence (siControl) from Ambion and using Hiperfect transfection reagent, for 48 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from each condition was extracted and purified using the RNA isolation kit (Macheray-Nagel).Qualitative analysis of total RNAs were performed in a bioanalizer Agilent 2100.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 100ng of total RNA using the LowInputQuick Amp Labeling kit – Agilent 5190-2305 according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
After fragmentation, 600 ng of labeled cRNA from each sample was hybridized in in situ hybridization oven (Agilent) for 17 h at 65ºC and washed during 1 min at room temperature in Gene Expression Wash Buffer 1 (Agilent) and 1 min at 37 ºC with Gene Expression Wahs buffer 2 (Agilent).
|
| Scan protocol |
Scanned on an Agilent G2539A scanner at 3um resolution and 100%PMT. The intensity data of each individual hybridization were extracted and the quality was assessed with the Feature Extraction software 10.7 (Agilent).
|
| Description |
M1 r1 +siC
|
| Data processing |
Scanned images and raw data were pre-processed with GeneSpring GX (Agilent).
|
| |
|
| Submission date |
Oct 14, 2016 |
| Last update date |
May 08, 2018 |
| Contact name |
Amaya Puig |
| E-mail(s) |
amaya.puig@salud.madrid.org
|
| Organization name |
Hospital Gregorio Marañón
|
| Department |
Immunology
|
| Lab |
Immunometabolism
|
| Street address |
Dr. Esquerdo 46
|
| City |
Madrid |
| State/province |
Madrid |
| ZIP/Postal code |
28007 |
| Country |
Spain |
| |
|
| Platform ID |
GPL21185 |
| Series (1) |
| GSE88768 |
Identification of PPARG regulated genes in human GM-CSF-primed monocyte-derived macrophages |
|
