title: Sample 6
organism: Mus musculus
description: Protocols: The F6 murine pro-B-cell leukaemia line was originally established from a spontaneous leukaemia in mice overexpressing IL-7 and has been previously described in Fisher, A. G., Burdet, C., Bunce, C., Merkenschlager, M. & Ceredig, R. Lymphoproliferative disorders in IL-7 transgenic mice: expansion of immature B cells which retain macrophage potential. Int. Immunol. 7, 415-423 (1995). F6 cells were grown in culture in IMDM with 5% FCS and were transduced with pRV-based retroviral vectors expressing either GFP or MHC II (mouse H2-Aa and H2-Ab1 chains). Retroviral particles were produced by transfection of pRV plasmids encoding MHC II chains or GFP into the Platinum-E (Plat-E) retroviral packaging cell line, using the GeneJuice transfection reagent (Novagen). Plat-E culture supernatants containing viral stocks were harvested two days after the initial transfection and used for transduction of target cells. Cells were mixed with supernatants containing viral stocks in the presence of 4 μg/ml polybrene (Sigma-Aldrich) and spun at ~315×g for 45 min, before resuspended in IMDM for further culture. F6 cells were grown in culture in IMDM with 5% FCS F6 cells were transduced either with GFP or with MHC II RNA was then extracted using the RNeasy Mini QIAcube or Micro kits (Qiagen) Illumina, RNA with PolyA selection (at GeneWiz)
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2018-08-07T17:02:06Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-05-15T14:09:59Z
attributes.tag: External Id
attributes.value: SAMEA4664368
attributes.tag: INSDC center name
attributes.value: Retroviral Immunology The Francis Crick Institute
attributes.tag: INSDC first public
attributes.value: 2018-08-07T17:02:06Z
attributes.tag: INSDC last update
attributes.value: 2018-05-15T14:09:59Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6763:Sample 6
attributes.tag: age
attributes.value: 6
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: F6
attributes.tag: cell type
attributes.value: pro-B cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: disease
attributes.value: pro-B cell leukaemia
attributes.tag: genotype
attributes.value: pRV-H2-Ab1
attributes.tag: organism part
attributes.value: lymph node
attributes.tag: phenotype
attributes.value: MHC II overexpression
attributes.tag: sample name
attributes.value: E-MTAB-6763:Sample 6
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57BL/6