Mus musculus
Protocols: Bone marrow was harvested from femur, tibia and hip bones in mice. Total bone marrow was depleted of differentiated hematopoietic cells (lineage-positive cells) using Mouse Hematopoietic Progenitor (Stem) Cell Enrichment Set (BD). Magnetically sorted Lineage-negative (lin-) cells were kept overnight at 4°C in IMDM medium supplemented with 10% FBS (HyClone) and 1% penicillin-streptomycin (Thermofisher). Staining was performed for 20min at room temperature (RT) using CD3ε (Lin) – APC clone 145-2C11 (553066, BD), TER-119 (Lin) – APC clone Ter-119 (557909, BD), CD45R/B220 (Lin) – APC  clone RA3-6B2 (553092, BD), Ly6G-6C (Lin)-APC clone RB6-8C5 (553129, BD), Ly-6A/E (Sca-1) - PeCy7 clone d7 (558162, BD), CD117 (c-Kit) – PE or PerCP-Cy5.5,  clone 2B8 (553355 or 560557 respectively, BD), CD34– FITC or AF700 clone RAM34 (560238 or 560518, BD), CD135 (Flk2) – BV421 or PE clone A2F10.1 (562898 or 553842 respectively, BD). Hematopoietic stem cells (Lin-Sca+c-Kit+CD34lowFlk2-) were sorted using ARIA3, ARIA Fusion or Influx cell sorters (BD Franklin Lakes, NJ, USA) and collected in Stem Span (StemCell).  Wild type (WT) C57BL/6J mice (2-4 months-old) are Tet2-/- littermates. All the mice were housed in a specific pathogen-free environment. All procedures were reviewed and approved by the Animal Care Committee (APAFIS #23286-2019121214392327 and #35147-2022020315419972). intraperitoneal injections of PBS or 6ug LPS were performed every two days for 30 days HSCs were lysed in Tri-Reagent (Zymo Research) and stored at -80°C until used. Total RNA was extracted using the Direct-Zol RNA microprep kit (Zymo research).  The RNA integrity and quantity was checked on the Agilent Fragment Analyzer (Agilent). The fragmentation time was adjusted according to RNA quality, after evaluation on the Agilent Fragment Analyzer. All samples were subjected to SMARTer cDNA synthesis using SMARTer Ultra Low Input RNA Kit for Sequencing - v3.   Double stranded cDNA was sheared using Covaris to obtain ds-cDNA in the 200–500 bp size range. ds-cDNA fragments were generated following the supplier protocole, amplified by PCR for 18 cycles and purified with AMPure XP beads (Beckman Coulter).
ENA first public
2026-07-28
INSDC center name
Inserm UMR1287, Université Paris-Saclay, Gustave Roussy, 94805, Villejuif, France.
INSDC status
public
Submitter Id
E-MTAB-15446:WT-LPS-3
age
2-4
broker name
ArrayExpress
cell type
hematopoietic stem cell
collection date
not collected
common name
house mouse
developmental stage
adult
genotype
wild type genotype
geographic location (country and/or sea)
not collected
immunophenotype
LSK FLK2- CD34-
isolate
not applicable
organism part
bone marrow
sample name
E-MTAB-15446:WT-LPS-3
scientific_name
Mus musculus
sex
female
strain
C57BL/6J