2025-08-13
ENA first public
ERC000011
ENA-CHECKLIST
German Cancer Research Centre (DKFZ)
INSDC center name
public
INSDC status
ERS25455979
SRA accession
E-MTAB-15477:Sample 1
Submitter Id
30
week
age
ArrayExpress
broker name
not collected
collection date
house mouse
common name
Protocols: Scl-tTA; H2B-GFP mice were treated with doxycycline for 100 days, which turn off constitutively expressed H2B-GFP, thus allow the marking of H2B-GFP+ dormant cells and H2B-GFP- active cell. Bone marrow was isolated after 100 days of chase, red cell lysed and lineage depleted using the EasySep Mouse Hematopoietic Progenitor Cell Isolation Kit (STEMCELL technologies, #19856) according to the manufacturer’s instruction. The Lin-depleted cells were then stained with the following antibodies: BV711 anti-mouse c-Kit (clone 2B8; BioLegend; 105835), APC-Cy7 anti-mouse Sca1 (clone D7; BD Biosciences; 560654), PE anti-mouse EPCR (clone eBio1560; eBioscience; 12-2012-82), eFluor 660 anti-mouse CD34 (clone RAM34; eBioscience; 50-0341-82), PE-Cy7 anti-mouse CD48 (clone HM48-1; BioLegend; 103424), BV421 anti-mouse CD135 (clone A2F10.1; BD Biosciences; 562898) or APC anti-mouse CD135 (clone A2F10; BioLegend; 135310),PE-Cy5 anti-mouse CD150 (clone TC15-12F12.2; BioLegend; 115912) or BV785 anti-mouse CD150 (clone TC15-12F12.2; BioLegend; 115937), Alexa Fluor 700 anti-mouse CD4 (clone GK1.5; eBioscience; 56-0041-82),Alexa Fluor 700 anti-mouse CD8a (clone 53-6.7; eBioscience; 56-0081-82), Alexa Fluor 700 anti-mouse CD11b (clone M1/70; eBioscience; 56-0112-82),Alexa Fluor 700 anti-mouse GR1 (clone RB6-8C5; eBioscience; 56-5931-82), Alexa Fluor 700 anti-mouse B220 (clone RA3-6B2; eBioscience; 56-0452-82), Alexa Fluor 700 anti-mouse Ter119 (clone TER-119; BioLegend; 116220). Index sorting was performed to sort single CD48– CD135- LSK cells into 384-well plate containing 2 µL RLP plus buffer (Qiagen, Cat #1053393) and 1 µL water per well and immediately frozen at -80 °C.  single cells were lysed in 2 µL RLP plus buffer (Qiagen, Cat #1053393) and 1 µL water per well and immediately frozen at -80 °C.  Single-cell methylome and transcriptome sequencing (scM&T-seq) libraries were generated based on the high-throughput protocol published previously (Cerrizuela et al., 2022) with the modification that no GpC methyltransferase incubation was done and that 384 cell could be multiplexed for the sequencing of the methylation layer.
description
adult
developmental stage
SCL-tTA; H2B-GFP
genotype
not collected
geographic location (country and/or sea)
CD48- CD135- LSK
immunophenotype
Pool of mice 1-3
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
bone marrow
organism part
Mus musculus
scientific_name
female
sex
SCL-tTA; H2B-GFP
strain
Sample 1
title