characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-27T00:21:11Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-27T00:21:11Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112329423
characteristics.characteristic: External Id
characteristics.text: EMBL
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-27T00:21:11Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-27T00:21:11Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14440333
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12609:BS_EMSeq_D14-5
characteristics.characteristic: Submitter Id
characteristics.text: 17
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: germinal centre B cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Mice were immunized intraperitoneally (i.p.) with 100μg of NP-CGG (4-hydroxy-3-nitrophenylacetyl-Chicken Gamma Globulin, Biosearch Technologies) dissolved in PBS and adsorbed on Alhydrogel aluminium hydroxide following manufacturer’s instructions (Invivogen). The impact of Hells loss-of-function on DNA methylation maintenance was quantified by enzymatic-methyl sequencing of sample from conditional knockout (Mb1-Cre+/WT HellsF/F) and littermate control mice (Mb1-CreWT/WT HellsF/F). Three time points were chosen to follow the kinetics: day 0 for naive B cells collection, day 7 and day 14 post NP-CGG intraperitoneal immunization for germinal center B cells collection. At each time point, 2 mice of each genotype were used. Live cells were FACS sorted (AriaIII) using surface markers as follows: B220+CD21+CD23+ for naive B cells, and B220+CD95+GL7+ for germinal center B cells. Due to their low frequency, germinal center B cells were FACS-sorted twice using the same gating strategy. Collected samples were lysed and processed for DNA extraction. DNA extraction from germinal centre B cells for EM-Seq was performed using the Kit Quick-DNA Microprep Plus (Zymo). DNA library construction protocol for EM-Seq, performed by EMBL GeneCore. The library was prepared according to NEBnext Enzymatic Methyl-seq kit (New England BioLabs, E7120).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: B220-positive; CD95-positive; GL7-positive
characteristics.characteristic: immunophenotype
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: spleen
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: BS_EMSeq_D14-5
characteristics.characteristic: title