Sample 24
Mus musculus
Protocols: Primary articular chondrocytes were isolated from five-day-old mice and cultured in DMEM/F12 supplemented with 10% FBS, Pen-Strep. To synchronise circadian rhythms, chondrocytes at 90% confluence in 35 mm cell culture dishes were incubated with 100 nM of dexamethasone for 60 min. Afterwards, media was changed and dishes were sealed with 40 mm glass cover slips. Sealed dishes were placed in an incubator at 37 °C for 112 hours. At 112 hours half the dishes (heat shock condition) were moved to an incubator at 43 °C for 60 min. The other dishes (dexamethasone condition) remained in an incubator at 37 °C and 100 nM dexamethasone were added to the culture media approximately 15 min prior to the end of the heat pulse. All dishes were then returned to the incubator at 37 °C and the first time-course sample was collected four hours later, after which chondrocytes were harvested for RNA isolation every four hours for 48 hours. This experiment was repeated three times utilising chondrocytes isolated from distinct litters. Total RNA was isolated using the PureLink RNA Mini Kit (Thermo Fisher) as per manufacturer's instructions. 50ng of mRNA per sample was used for library generation. Libraries were generated using the TruSeq Stranded mRNA assay (Illumina) according to the manufacturer's protocol.
organism
Mus musculus
organism
Mus musculus
collection date
not collected
stimulus
none
scientific_name
Mus musculus
cell type
chondrocyte
common name
house mouse
organism part
cartilage
developmental stage
postnatal
genotype
wild type genotype
isolate
not applicable
geographic location (country and/or sea)
not collected