|
Status |
Public on Dec 01, 2014 |
Title |
Macrophage Donor 1 day 3 [5 Hy5TM] |
Sample type |
RNA |
|
|
Source name |
Monocyte-derived macrophages
|
Organism |
Homo sapiens |
Characteristics |
cell type: Macrophage donor: 1 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
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|
|
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. |
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