Sample 3
Homo sapiens
Protocols: Briefly, U87MG-GFP cells were implanted intracranially into striatum of male NOD/SCID mice (6-8 weeks) and allowed to grow for three weeks. Hoechst 33342 (vascular perfusion dye) was injected as onto the tail vein and the tumor resected post 5 min incubation. Tumors were isolated, immediately dissociated into single cells. Single cell suspensions were sorted using BD FACSAria™ III. The gating strategy was used to isolate only GFP+ cancer cells (to exclude all stromal elements) and these cells were further fractionated into two fractions - Hh (Hoechst High) and Hl (Hoechst low) based on their dye uptake. Hh - are the cells closest to blood vasculature and Hl are the farthest. The sorted cells (~50000 cells) were immediately centrifuged, supernatant removed and resuspended in 500 µl of Trizol. RNAs were isolated from the respective sorted fractions using RNeasy mini kit (Qiagen, 74004). RNA quality was determined with the aid of a TapeStation (Agilent Technologies). Libraries were prepared and sequenced using Illumina directional RNA sequencing protocol (Mi-Seq Reagent Kit V3).
ENA-FIRST-PUBLIC
2019-04-30T04:02:49Z
ENA-LAST-UPDATE
2019-03-19T10:54:50Z
External Id
SAMEA5522490
INSDC center name
HEBREW UNIVERSITY OF JERUSALEM
INSDC first public
2019-04-30T04:02:49Z
INSDC last update
2019-03-19T10:54:50Z
INSDC status
public
Submitter Id
E-MTAB-6881:Sample 3
broker name
ArrayExpress
cell line
U-87 MG
common name
human
disease
glioblastoma multiforme
genotype
GFP expression under CMV promoter
organism part
brain
phenotype
Hoechst low
proximity to blood vessels
far
sample name
E-MTAB-6881:Sample 3
scientific_name
Homo sapiens
strain
NOD-SCID
xenograft
intracranial