|
| Status |
Public on Dec 01, 2014 |
| Title |
Macrophage Donor 3 day 3 [11 Hy5TM] |
| Sample type |
RNA |
| |
|
| Source name |
Monocyte-derived macrophages & Monocyte-derived osteoclasts
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Macrophage
donor: 3
time point: day 3
|
| Treatment protocol |
Cells were left untreated.
|
| Growth protocol |
CD14+ monocytes were cultured at 4×106 cells/ml in: (A, osteoclasts) RPMI 1640 medium supplemented with 10% FBS, 2 mmol/L L-glutamine, penicillin (100 U/ml), streptomycin (100 μg/ml), 33ng/ml M-CSF, 100ng/ml sRANKL, or (B, macrophages) RPMI 1640 medium supplemented with 10% FBS, 2 mmol/L L-glutamine, penicillin (100 U/ml), streptomycin (100 μg/ml), 33ng/ml M-CSF. Media was replenished every 72 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the miRNeasy kit (Qiagen) following manufacturer’s protocol. The RNA was quantitated using the NanoDrop (Thermo Scientific) and integrity assessed using the 2100 Bioanalyzer.
|
| Label |
Hy5TM
|
| Label protocol |
250ng of total RNA was used as input for miRCURY LNATM microRNA Hi-Power Labeling Kit. Labelling was performed on an automated Tecan hybridization station.
|
| |
|
| Hybridization protocol |
Hybridisation was performed by Exiqon.
|
| Scan protocol |
Scanning was performed by Exiqon.
|
| Description |
The samples were labeled using the miRCURY LNATM microRNA Hi-Power Labeling Kit, Hy3TM/Hy5TM and hybridized on the miRCURY LNATM microRNA Array (7th Gen)
|
| Data processing |
Data normalization was performed using Quantile normalization by Exiqon.
Fold change (Normalized Reads)
Expression analysis (ANOVA for Time - Macrophages)
Expression analysis (ANOVA for Time – Macrophages, excluding monocytes)
Expression analysis (ANOVA for Time - Osteoclasts)
Expression analysis (ANOVA for Time – Osteoclasts, excluding monocytes)
Expression analysis for Mac_1 vs. Ost_1
Expression analysis for Mac_3 vs. Ost_3
Expression analysis for Mac_6 vs. Ost_6
Expression analysis for Mac_12 vs. Ost_12
|
| |
|
| Submission date |
Aug 27, 2014 |
| Last update date |
Dec 01, 2014 |
| Contact name |
Jezrom Bokcaerin Self-Fordham |
| E-mail(s) |
jez.fordham@gmail.com
|
| Phone |
7735247565
|
| Organization name |
UIC
|
| Department |
Periodontics
|
| Lab |
461
|
| Street address |
801 S Paulina
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60612 |
| Country |
USA |
| |
|
| Platform ID |
GPL19128 |
| Series (1) |
| GSE60840 |
miRCURY LNATM microRNA Array (7th Gen) profiling of monocyte-derived osteoclasts and macrophages. |
|
