characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-10-27T00:39:09Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-10-27T00:39:09Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA113400309
characteristics.characteristic: External Id
characteristics.text: umea university, dept molecular biology
characteristics.characteristic: INSDC center name
characteristics.text: 2023-10-27T00:39:09Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-10-27T00:39:09Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS15395456
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12902:samp126
characteristics.characteristic: Submitter Id
characteristics.text: 1 to 4
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: microglial cell
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: Cortical astrocytes and neurons were generated. In brief, astrocytes were isolated from neonatal mice between postnatal day 1 and 4. Cerebral cortices were isolated in HBSS (Gibco) containing 0.5% penicillin and streptomycin (Gibco) and 0.4% glucose. Tissues were dissociated by trituration and seeded in poly-d-lysine-coated T-75 tissue culture flasks. Astrocytes were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat-inactivated FBS (Thermo Fisher Scientific), 0.2% penicillin and streptomycin (Gibco) and 2 mM L-glutamine (Gibco). Primary cortical neurons were isolated from the cerebral cortices at embryonic day 17 and seeded onto poly-d-lysine-coated wells. Neurons were grown in neurobasal plus medium (Gibco) supplemented with B27 plus supplement (Gibco), 0.2% penicillin and streptomycin. Microglia were isolated from the astrocyte cultures as previously described [23] and grown in (DMEM) supplemented with 10% heat-inactivated FBS (Thermo Fisher Scientific), 0.2% penicillin and streptomycin (Gibco) and 2 mM L-glutamine (Gibco) and conditioned astrocytes medium (1:1). Monolayers of neurons (MOI 0.01), astrocytes (MOI 0.1) and microglia (MOI 1) were infected with LGTV or TBEV for 1 h at 37˚C and 5% CO2 before the inoculum was removed and replaced with fresh medium. Cell supernatant was harvested at the indicated time points and viral titers were determined by focus forming assay. Total RNA from infected or mock-treated monocultures was extracted using Nucleospin RNA plus mini kit (Macherey-Nagel) according to manufacturer’s instructions. The Smart-Seq2 protocol was used to produce fragmented cDNA with Nextera sequencing adapters.
characteristics.characteristic: description
characteristics.text: juvenile stage
characteristics.characteristic: developmental stage
characteristics.text: IFNAR -/-
characteristics.characteristic: genotype
characteristics.text: nunya43
characteristics.characteristic: nunya_wellid
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: brain
characteristics.characteristic: organism part
characteristics.text: 94.387
characteristics.characteristic: rna concentration
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: mixed
characteristics.characteristic: sex
characteristics.text: C57BL/6J
characteristics.characteristic: strain
characteristics.text: samp126
characteristics.characteristic: title