|
| Status |
Public on May 02, 2014 |
| Title |
human_monocytes_0day in DC culture |
| Sample type |
RNA |
| |
|
| Source name |
human monocyte derived cells
|
| Organism |
Homo sapiens |
| Characteristics |
in vitro culture: 0day in GMCSF/IL4 DC culture
|
| Treatment protocol |
DCs were matured by 300ng/ml LPS (0111:B4, Sigma) on day 5 and harvested on day 7 for mature DCs.
|
| Growth protocol |
Monocytes were purified from PBMCs using anti-CD14 microbeads (Miltenyi Biotech) and then cultured at 37°C in 24-well plates (5x10^5 cells per well) in 1 mL of RPMI-1640 Medium (PAA) with 10% (vol/vol) FCS (PAA) containing 100 ng/mL human GM-CSF and 20 ng/mL human IL-4 (R&D Systems，Minneapolis, MN). Half medium was replaced by fresh medium with GM-CSF and IL-4 at day 3 and day 5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cells at indicated time point with Trizol.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cDNA were prepared according to the standard Affymetrix protocol from 250 ng total RNA by using Ambion® WT Expression Kit. WT Expression Kit
|
| |
|
| Hybridization protocol |
Following labeling, 5.5 ug of cDNA were hybridized for 16 hr at 45℃ on GeneChip Human Transcriptome Array 2.0 in Hybridization Oven 645. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scaned by using Affymetrix® GeneChip Command Console (AGCC) which installed in GeneChip® Scanner 3000 7G.
|
| Data processing |
The data were analyzed with Robust Multichip Analysis (RMA) algorithm using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
Jan 16, 2014 |
| Last update date |
May 02, 2014 |
| Contact name |
PIN WANG |
| E-mail(s) |
pinqi21sh@163.com
|
| Organization name |
SMMU
|
| Street address |
Xiangyin Road 800
|
| City |
Shanghai |
| ZIP/Postal code |
200433 |
| Country |
China |
| |
|
| Platform ID |
GPL17586 |
| Series (1) |
| GSE54143 |
DC-specific lincRNA controls dendritic cell development and function through interacting with STAT3 |
|
