PRJNA471582: Genetic and transcriptional variation alters cancer cell line drug response [MCF7 strain AA]
Summary: 10X Genomics single cell RNAseq of MCF7 cells treated with bortezomib.Human cancer cell lines are the workhorse of cancer research. While cell lines are known to evolve in culture, the extent of the
Overall Design: MCF7 cells (strain AA) were treated with bortezomib (500nM) and harvested before treatment, after 12 hours of exposure (t12), after 24 hours of exposure (t48), or after 72 hours of exposure followed by drug wash and 24 hours of recovery (t72+24)
||MCF7 cells were cultured in RPMI-1640 (Life Technologies), with 10% Fetal Bovine Serum (Sigma-Aldrich) and 1% Penicillin-Streptomycin-Glutamine (Life Technologies).
||MCF7-AA cells were exposed to 500nM of bortezomib (Selleckchem S1013 ) and harvested before treatment, after 12 hours of exposure (t12), after 24 hours of exposure (t48), or after 72 hours of exposure followed by drug wash and 24 hours of recovery (t72+24).
||DissociateCells were washed, trypsinized, passed through a 40μM cell strainer, centrifuged at 400g, and resuspended at a concentration of 1,000 cells/μL in PBS + 0.5% BSA.Single cells were processed through the Chromium Single Cell 3′ Solution platform using the Chromium Single Cell 3’ Gel Bead, Chip and Library Kits (10X Genomics), as per the manufacturer’s protocol.
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||Illumina HiSeq 2500
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