characteristics.text: 2016-04-30T17:09:59Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:19:32Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2056773
characteristics.characteristic: External Id
characteristics.text: EMBL-EBI
characteristics.characteristic: INSDC center name
characteristics.text: 2016-04-30T17:09:59Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:19:32Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS221079
characteristics.characteristic: SRA accession
characteristics.text: E-SYBR-2:DC DC6704 untreated
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: human
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: common name
characteristics.text: Protocols: Fungal cell preparation: 5 S. cerevisiae strains were grown at 28degC with stirring (150 r.p.m.) until mid-exponential phase in liquid YPD medium (1%, w/v yeast extract, 2%, w/v peptone and 2% w/v glucose). 5 C. albicans and parapsilosis strains were grown at at 28degC with stirring (150 r.p.m.) until mid-exponential phase in liquid Saboraud medium (1%, w/v peptone and 2% w/v glucose). Cells were then harvested by centrifugation, washed twice with sterile water and resuspended in fresh RPMI 1640 complete medium. 4 A. fumigatus strains were grown at 37degC in solid Saboraud medium. Spores were then collected using PBS + 0.1% Tween and then washed twice with sterile water and resuspended in fresh RPMI 1640 complete. Human cell preparation: PBMCs were isolated from buffy coat blood sample from 6 healthy donors from the Transfusion Unit of the Careggi Hospital (Florence, Italy) by Ficoll-Hypaque density gradient centrifugation (Biochrom AG). Monocytes were isolated from low density PBMCs by magnetic enrichment with anti-CD14 beads (Miltenyi Biotec). Cells were cultured in RPMI complete medium (RPMI 1640, 10% w/v FBS, 1% w/v 200 mM glutamine, 1% w/v 50 mM sodium pyruvate, penicillin-streptomicin) in the presence of GM-CSF (800 U/ml) and recombinant IL-4 (1000 U/ml) for 6 days to allow dendritic cell (DC) differentiation (37degC, 5% CO2). Cell treatment: DCs were cultured for 4 hours with the different fungal strains at a stimuli:DC ratio of 5:1 (time= 4hr, 37degC, 5% CO2, complete RPMI medium). As controls, DCs or the single fungal strain were grown alone in the same experimental condition. RNA was extracted from cells using the standard TriZol protocol. For A. fumigatus containing samples, prior to TriZol extraction, fungal cells were disrupted by grinding in liquid nitrogen with pestle and mortar. RNA samples were then quantified, and their integrity checked using NanoChip BioAnalyzer technology (Agilent). For each sample, one paired-end mRNA library with approximately 150bp insert size was prepared using the TruSeq RNA Sample Preparation Kit v2 (Illumina).
characteristics.characteristic: description
characteristics.text: DC DC6704
characteristics.characteristic: individual
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9606
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: DC DC6704 untreated
characteristics.characteristic: title