|
| Status |
Public on Jan 25, 2019 |
| Title |
Primary macrophages 2hr stimulation with LPS |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Cells stimulated with LPS for 2hrs (pooled from 4 donors)
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Monocyte-derived macrophages
stimulator: LPS
time: 2 hours
|
| Treatment protocol |
Cells were plated at 1x10e6 cells/ml. The next day cells were stimulated with 100ng/ml TLR-grade LPS and/or 10ng/ml IL-10 for the indicated time. Cells were washed once with 1xPBS and harvested with the MiRvana extraction/homogenisation buffer
|
| Growth protocol |
Primary human monocytes were isolated by elutriation from plateletphoresis residues (blood cones) and differentiated using 10ng/ml M-CSF for 4 days
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using the miRvana kit (Ambion)
|
| Label |
Hy3
|
| Label protocol |
750ng total RNA was mixed with miRCURY LNA™ microRNA Array Spike-in controls (#208040) and then labelled with the miRCURY LNA™ microRNA Power labeling kit (#208032-A) for dual labelling with Hy3 (samples) and Hy5 (common reference) according to manufacturers recommendations
|
| |
|
| Channel 2 |
| Source name |
Common Reference
|
| Organism |
Homo sapiens |
| Characteristics |
preparation: Equal quantities of RNA from each of the six experimental data points
|
| Treatment protocol |
Cells were plated at 1x10e6 cells/ml. The next day cells were stimulated with 100ng/ml TLR-grade LPS and/or 10ng/ml IL-10 for the indicated time. Cells were washed once with 1xPBS and harvested with the MiRvana extraction/homogenisation buffer
|
| Growth protocol |
Primary human monocytes were isolated by elutriation from plateletphoresis residues (blood cones) and differentiated using 10ng/ml M-CSF for 4 days
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using the miRvana kit (Ambion)
|
| Label |
Hy5
|
| Label protocol |
750ng total RNA was mixed with miRCURY LNA™ microRNA Array Spike-in controls (#208040) and then labelled with the miRCURY LNA™ microRNA Power labeling kit (#208032-A) for dual labelling with Hy3 (samples) and Hy5 (common reference) according to manufacturers recommendations
|
| |
|
| |
| Hybridization protocol |
Hybridisation took place in miRCURY LNA™ microRNA Array Hybridization buffer (#208022) and washing was with the miRCURY LNA™ microRNA Array, Wash buffer kit (#208021) following the manufacturer's reccomendations
|
| Scan protocol |
not available
|
| Description |
2hrs LPS compared to the common reference (pooled donors)
|
| Data processing |
Background subtraction used Normexp with offset value 1 (convolution model described by Ritchie ME, Bioinformatics 2007: 23 p2700-7 ) Normalisation was by LOWESS regression. Spots where no signal above background was detected were removed
|
| |
|
| Submission date |
Jan 24, 2019 |
| Last update date |
Jan 25, 2019 |
| Contact name |
RACHEL SIMMONDS |
| E-mail(s) |
rachel.simmonds@surrey.ac.uk
|
| Phone |
01483684714
|
| Organization name |
University of Surrey
|
| Street address |
FHMS UNIVERSITY OF SURREY, STAG HILL CAMPUS
|
| City |
GULDFORD |
| State/province |
Surrey |
| ZIP/Postal code |
GU2 7XH |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL7722 |
| Series (1) |
| GSE125572 |
microRNA responses of LPS and IL-10 stimulated primary human monocyte-derived macrophages over a short time-course |
|
