title: 6B
organism: Mus musculus
description: Protocols: RAW264.7 cells were cultured for 48 h in 6-well plates (Greiner) in phenol red-free alpha MEM supplemented with 10% dextran-coated charcoal-stripped FBS with ultra-low endotoxin levels (Biosera). Cells were treated with vehicle (0.1% DMSO), dexamethasone (1 μM) or AZD9567 (1 μΜ) in the presence or absence of lipopolysaccharide (LPS, 10 ng/ml) for 1 h. At the end of the treatment cells were collected in TRIzol Reagent and used for RNA extraction. RNA was extracted from cells using the TRIzol Reagent (Invitrogen) followed by a clean-up step combined with on-column DNase digestion using the RNeasy kit (Qiagen) according to manufacturers&#39; instructions. After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT) are used to isolate mRNA. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments are selected for the PCR amplification as templates.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2023-06-30T00:24:23Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2023-06-30T00:24:23Z
attributes.tag: External Id
attributes.value: SAMEA112864441
attributes.tag: INSDC center alias
attributes.value: Institute of Chemical Biology, National Hellenic Research Foundation
attributes.tag: INSDC center name
attributes.value: Institute of Chemical Biology, National Hellenic Research Foundation
attributes.tag: INSDC first public
attributes.value: 2023-06-30T00:24:23Z
attributes.tag: INSDC last update
attributes.value: 2023-06-30T00:24:23Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-12816:6B
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: RAW 264.7
attributes.tag: common name
attributes.value: house mouse
attributes.tag: disease
attributes.value: normal
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: ascitic fluid
attributes.tag: sample name
attributes.value: E-MTAB-12816:6B
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: strain
attributes.value: BALB/c