GFP-WT
Homo sapiens
Protocols: Cells were transfected using lentiviral pV81 vectors with doxycycline inducible expression of WT or mutant SPI1 fused to GFP (pV81/GFP-SPI1) or GFP alone. The pV81 backbone was described in this reference : Calvo J, BenYoucef A, Baijer J, Rouyez MC, Pflumio F. Assessment of human multipotent hematopoietic stem/progenitor cell potential using a single in vitro screening system. PLoS One. 2012;7:e50495. Tumor cells were defined by the positivity of CD19 and monotypic light chain (LC) while control samples were CD3-positive T lymphocytes. The cellular populations were sorted using an Influx cell sorter (BD Biosciences, San Jose, CA, USA). The sorted cells were subjected to an additional round of sorting to provide a cell fraction purity of more than 98% and subjected to DNA/RNA extraction using the AllPrep DNA/RNA Kit (Qiagen). Sheared chromatin was immunoprecipitated using the following antibodies: anti-SPI1 (Santa-Cruz, sc-352, clone T21), anti-GFP-Trap (Chromotek) and rabbit IgG (Santa-Cruz, sc-2027). Tumor cells were defined by the positivity of CD19 and monotypic light chain (LC) while control samples were CD3-positive T lymphocytes. The cellular populations were sorted using an Influx cell sorter (BD Biosciences, San Jose, CA, USA). The sorted cells were subjected to an additional round of sorting to provide a cell fraction purity of more than 98% and subjected to DNA/RNA extraction using the AllPrep DNA/RNA Kit (Qiagen). Enriched DNA from ChIP and Input DNA fragments were end-repaired, extended with an 'A' base on the 3′end, ligated with indexed paired-end adaptors (NEXTflex, Bioo Scientific, Proteigene, Saint Marcel, Fra nce) using the Bravo Platform (Agilent, Les Ulis, France), size-selected after 4 cycles of PCR with AMPure XP beads (Beckman Coulter, Villepinte, France) and amplified by PCR for 10 cycles more.
ENA-FIRST-PUBLIC
2019-05-01T04:02:54Z
ENA-LAST-UPDATE
2019-04-16T15:16:16Z
External Id
SAMEA5568575
INSDC center name
INSERM U1170, Gustave Roussy, Villejuif, France
INSDC first public
2019-05-01T04:02:54Z
INSDC last update
2019-04-16T15:16:16Z
INSDC status
public
Submitter Id
E-MTAB-7881:GFP-WT
broker name
ArrayExpress
cell line
OCI-LY10
cell type
lymphoma B cell
common name
human
disease
diffuse large B-cell lymphoma
genotype
MYD88-LP; pV81-GFP
organism part
blood
sample name
E-MTAB-7881:GFP-WT
scientific_name
Homo sapiens