Day_14_DTR_neg
Protocols: Tumors were collected, weighed and minced into small pieces. To isolate naive flank skin, hair was shaved, adipose tissue was removed and skin tissue was minced. All tissues were subsequently enzymatically digested using a cocktail of dispase (Life Technologies, Carlsbad, CA), collagenase P and DNaseI (Roche, Penzberg, Germany) for 45 min at 37°C, to obtain a single cell suspension. Cells were counted using a Vi-CELL XR (Beckman Coulter, Brea, CA). For cytokine staining, cell suspensions were stimulated with eBioscience Cell Stimulation Cocktail plus protein transport inhibitors (cat. 00-4975-93) resuspended in RPMI with 10% FBS plus 2 mmol/L l-glutamine and 2-Mercaptoethanol for 2 hours at 37°C.Cells were labelled with monoclonal antibodies purchased from BioLegend. Samples were enriched for CD24-CD45- stromal cells. Sorted cells were loaded onto a 10x Chromium Chip using reagents from the Chromium Single Cell 3' Library Kit (10x Genomics) according to the manufacturer's protocol. Amplified cDNA was used for 3' RNA-seq library generation, RNA-seq libraries were prepared following the manufacturer's user guide (10x Genomics).
ENA-FIRST-PUBLIC
2022-08-17
ENA-LAST-UPDATE
2022-08-17
External Id
SAMEA110471496
INSDC center alias
GENENTECH
INSDC center name
GENENTECH
INSDC first public
2022-08-17T00:15:15Z
INSDC last update
2022-08-17T00:15:15Z
INSDC status
public
Submitter Id
E-MTAB-12036:Day_14_DTR_neg
broker name
ArrayExpress
cell_type
fibroblast
common name
house mouse
developmental stage
adult
disease
cancer
genotype
wild type genotype
growth condition
injected subcutaneous with KPR cells
immunophenotype
CD24-CD45-
organism part
skin of body
sample name
E-MTAB-12036:Day_14_DTR_neg