description: Protocols: Bone-marrow cells from naïve mice were isolated by flushing femurs, tibias and humeri with RPMI-1640 medium. Following suspended in ACK lysis buffer to lyse red blood cells, bone marrow cells were passed through a 40 µm filter. Then cells were resuspended and cultured in complete RPMI-1640 medium containing GM-CSF and IL-4. The cultured medium was entirely discarded on day 4 and new warmed medium supplemented with GM-CSF and IL-4 was added. On day 7, the cultured cells were collected and used for further experiments. BM-DCs were incubated with WT or Ptgs2-/- lung fibroblast-derived conditioned medium or purified PGE2. Total RNA was isolated from BM-DCs using the miRNeasy Mini kit (Qiagen). Stranded libraries were prepared using the KAPA mRNA HyperPrep Kit: isolation of polyA containing mRNA using oligo-dT magnetic beads, RNA fragmentation, first and second strand cDNA synthesis, ligation of Illumina-specific adapters containing a unique barcode sequence for each library, and PCR amplification. Libraries were checked for quality and concentration using the DNA 1000 assay and quantitative PCR, following the manufacturers' instructions.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-07-19
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-07-19
attributes.tag: External Id
attributes.value: SAMEA110186531
attributes.tag: INSDC center alias
attributes.value: THE JACKSON LABORATORY
attributes.tag: INSDC center name
attributes.value: THE JACKSON LABORATORY
attributes.tag: INSDC first public
attributes.value: 2022-07-19T00:15:36Z
attributes.tag: INSDC last update
attributes.value: 2022-07-19T00:15:36Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11873:Sample 6
attributes.tag: age
attributes.value: 10
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_type
attributes.value: dendritic cell
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: growth condition
attributes.value: co-culture with Ptgs2 -/- lung fibroblasts
attributes.tag: individual
attributes.value: 6
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: sample name
attributes.value: E-MTAB-11873:Sample 6
attributes.tag: sex
attributes.value: female
attributes.tag: stimulus
attributes.value: none
attributes.tag: strain
attributes.value: C57BL/6J