|
| Status |
Public on Oct 06, 2021 |
| Title |
MH-S_IL-4 rep3 |
| Sample type |
RNA |
| |
|
| Source name |
alveolar macrophage
|
| Organism |
Mus musculus |
| Characteristics |
treatment: IL-4, 48h
cell: alveolar macrophage
|
| Treatment protocol |
MH-S cells were seeded into plastic 6-well plates at a concentration of 1×105 cells/mL and incubated in serum-deprived medium for 12 h followed by treatment with IL-4 at 20 ng/mL for 48 h.
|
| Growth protocol |
The murine alveolar macrophages (MH-S) purchased from ATCC were maintained in RPMI 1640 containing 10% FBS supplemented with 1% penicillin/streptomycin and incubated at 37 °C in a humidified incubator containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol (Invitrogen) and purified with RNeasy mini kit (QIAGEN) according to the manufacturer’s instructions. RNA quality and quantity was measured by using nanodrop spectrophotometer (ND-1000, Nanodrop Technologies) and RNA Integrity was determined by gel electrophoresis.
|
| Label |
Hy3
|
| Label protocol |
The miRCURY™ Hy3™/Hy5™ Power labeling kit (Exiqon, Vedbaek, Denmark) was used according to the manufacturer's guideline for miRNA labelling.
|
| |
|
| Hybridization protocol |
The Hy3™-labeled samples were hybridized on the miRCURYTM LNA Array (v.19.0) (Exiqon) according to array manual.
|
| Scan protocol |
The slides were scanned using the Axon GenePix 4000B microarray scanner (Axon Instruments, Foster City, CA).
|
| Description |
miRNA expression after 48hr in lL-4-induced alveolar macrophages
|
| Data processing |
Scanned images were then imported into GenePix Pro 6.0 software (Axon) for grid alignment and data extraction. Replicated miRNAs were averaged and miRNAs that intensities>=30 in all samples were chosen for calculating normalization factor. Expressed data were normalized using the Median normalization. After normalization, significant differentially expressed miRNAs between two groups were identified through Fold change and P-value. Differentially expressed miRNAs between two samples were filtered through Fold change. Finally, hierarchical clustering was performed to show distinguishable miRNA expression profiling among samples.
|
| |
|
| Submission date |
Jan 15, 2020 |
| Last update date |
Oct 06, 2021 |
| Contact name |
Qiujie Wang |
| E-mail(s) |
wangqiuj@mail2.sysu.edu.cn
|
| Organization name |
Sun Yat-sen University Second University Hospital
|
| Department |
Department of Respiratory Medicine
|
| Street address |
No.107 Yan jiang West Road
|
| City |
Guangzhou |
| State/province |
Guangdong |
| ZIP/Postal code |
510120 |
| Country |
China |
| |
|
| Platform ID |
GPL18058 |
| Series (1) |
| GSE143740 |
miRNA expression profiling and identification of a miRNA regulator for macrophage polarization in the murine alveolar macrophage cell line MH-S. |
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