Sample 2
Mus musculus
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)
ENA-FIRST-PUBLIC
2018-08-07T17:02:06Z
ENA-LAST-UPDATE
2018-05-15T14:09:59Z
External Id
SAMEA4664364
INSDC center name
Retroviral Immunology The Francis Crick Institute
INSDC first public
2018-08-07T17:02:06Z
INSDC last update
2018-05-15T14:09:59Z
INSDC status
public
Submitter Id
E-MTAB-6763:Sample 2
age
6
broker name
ArrayExpress
cell line
F6
cell type
pro-B cell
common name
house mouse
disease
pro-B cell leukaemia
genotype
pRV-H2-Aa
organism part
lymph node
phenotype
empty retroviral vector
sample name
E-MTAB-6763:Sample 2
scientific_name
Mus musculus
sex
male
strain
C57BL/6