PMC:2685087 / 8317-9376 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/2685087","sourcedb":"PMC","sourceid":"2685087","source_url":"https://www.ncbi.nlm.nih.gov/pmc/2685087","text":"Cell culture and transfections\nMCF7, T47D, MELN (28) and COS-1 cells were maintained in DMEM supplemented with 10% DSS, 1% penicillin/streptomycin and 2% glutamine. HeLa cell line expressing wild-type ERα has been described elsewhere (29) (a kind gift from Dr Shapiro). All cells were incubated at 37°C in a humidified 5% CO2 atmosphere. MCF7 and MELN cells were transfected with 5 nM siRNA using Hiperfect according to the manufacturer's instructions (Qiagen, Crawley, UK). Transient transfections of COS-1 and MCF7 were performed 24 h after seeding cells using FuGENE 6 according to the manufacturer's instructions (Roche, Sussex, UK). Cells were maintained in phenol red-free media with 5% charcoal-stripped serum 48 h before experimentation. The CK1δ isoform was silenced using two independent siRNAs, targeting the following sequences: 5′-ccggtct aggatcgaaatgtt-3′ and 5′-ctccctgacgattccactgta-3′. At 48 h (or 72 h) post-transfection, cells were treated with either vehicle (ethanol) or E2, and harvested later (as indicated) for RNA or protein analyses.","divisions":[{"label":"Title","span":{"begin":0,"end":30}}],"tracks":[{"project":"2_test","denotations":[{"id":"19339517-10492897-76537767","span":{"begin":49,"end":51},"obj":"10492897"},{"id":"19339517-17615152-76537768","span":{"begin":235,"end":237},"obj":"17615152"}],"attributes":[{"subj":"19339517-10492897-76537767","pred":"source","obj":"2_test"},{"subj":"19339517-17615152-76537768","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#93e1ec","default":true}]}]}}