|
Status |
Public on Jan 01, 2020 |
Title |
CD44KO-Con |
Sample type |
SRA |
|
|
Source name |
Hepatic stellate cell
|
Organism |
Mus musculus |
Characteristics |
strain: C57BL/6 cell: Primary hepatic stellate cell genotype: CD44-/-
|
Treatment protocol |
24-week-old male wild-type, TLR4-/-, and CD44-/- mice were used. All procedures were approvd by the Institutional Animal Care and Use Committee at Cedars-Sinai Medical Center. Hepatic stellate cells were isolated from the mice in a 2-step process: collagenase-pronase perfusion of mouse livers followed by Nycodenz 2-layer discontinuous density gradient centrifugation. We further depleted contaminated liver macrophages by magnetic antibody cell sorting (Miltenyi Biotec) using Cd11b-conjugated microbeads (Miltenyi Biotec).
|
Growth protocol |
Primary hepatic stellate cells were isolated from male wild-type, TLR4-/-, and CD44-/- mice. After, the isolated primary hepatic stellate cells were incubated in DMEM medium containing 10% FBS, and supplemented with Antibiotic-Antimycotic for 3 days, serum was deprived for 24 hr.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted using TRIzol (Invitrogen) and purified using RNA isolation kit (Macherey-Nagel). Beijing Genomics Institute (BG) (Beijing, China) performed the RNA sequencing. RNA libraries were prepared for sequencing using standard BGISEQ-500 protocols
|
|
|
Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
BGISEQ-500 |
|
|
Data processing |
Sequenced reads were trimmed for adaptor sequence, and masked for low-complexity or low-quality sequence, then mapped to mm10 whole genome using version 2.2.5 with parameters--phred64 --sensitive --dpad 0 --gbar 99999999 --mp 1,1 --np 1 --score-min L,0,-0.1 -p 16 -k 200 Fragments Per Kilobase Million(FPKM) were calculated using RSEM Genome_build: mm10 Supplementary_files_format_and_content: Excel files include FPKM values for each Sample.
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|
|
Submission date |
Jan 15, 2018 |
Last update date |
Jan 01, 2020 |
Contact name |
Yoon Mee Yang |
E-mail(s) |
[email protected]
|
Organization name |
Kangwon National University
|
Department |
Pharmacy
|
Street address |
1 Kangwondaehak-gil
|
City |
Chuncheon-si |
State/province |
Gangwon-do |
ZIP/Postal code |
24341 |
Country |
South Korea |
|
|
Platform ID |
GPL23479 |
Series (1) |
GSE109200 |
TLR4 or CD44 deficiency effect on hepatic stellate cells |
|
Relations |
BioSample |
SAMN08366018 |
SRA |
SRX3584898 |