characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-21T10:00:07Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-21T10:00:07Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112330147
characteristics.characteristic: External Id
characteristics.text: BIMSB
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-21T10:00:07Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-21T10:00:07Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14441057
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12522:R98AC99A_br3
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: B cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: RNA prepared from isolated murine splenic B cells transduced with MIG control virus or IRF4-WT, IRF4-C99R, or IRF4-R98AC99A .Splenic B cells were isolated from C57BL/6 mice by CD43 depletion with magnetic anti-mouse CD43 microbeads (Miltenyi Biotech, Cat#130-049-801) according to the manufacturer’s instructions. To induce B cell activation and terminal differentiation, purified B cells (density: 1x106 cells/mL; 4x106 cells per well) were cultured in the presence of recombinant mouse IL-4 (25ng/mL; R&D; Cat#404-ML) and LPS (20µg/mL; Sigma; Cat#L2880) over night. 24h after isolation, B cells were collected (300xg, 5min, 4°C) and resuspended in B cell medium supplemented with 8µg/mL polybrene (EMD Millipore, #TR-1003) at a density of 2x106 cells/mL. To introduce IRF4 wildtype and mutant, 4x106 B cells per well were plated in 2mL on 6-well plates that had been coated with RetroNectin (Takara, #T100B; 25µg/mL; 4°C, overnight), blocked with 2% BSA (in PBS, 1h, RT) and pre-loaded with the respective retroviral particles (1h, 37°C). Retroviral transduction was performed by the addition of 2mL of the respective retroviral supernatant and subsequent centrifugation (800xg, 90min, 32°C). 24h after transduction, B cells were collected (300xg, 5min, 4°C), resupended in new B cell medium and cultured (density: 1x106 cells/mL; 4x106 cells per well) for another 72h (FACS for RNA-seq, flow cytometric analysis of plasma cell differentiation) in the presence of recombinant mouse IL-4 (25ng/mL; R&D; Cat#404-ML) and LPS (20µg/mL; Sigma; Cat#L2880). Nucleic acids were extracted with RNeasy Micro Kit (Qiagen) per provider's protocol Nucleic acid libraries were constructed as described in the accompanying manuscript
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: IRF4 R98AC99A mutant
characteristics.characteristic: genotype
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: C57BL/6J
characteristics.characteristic: strain
characteristics.text: R98AC99A_br3
characteristics.characteristic: title