AR pro, Palbociclib treatment
Homo sapiens
Protocols: Cells were maintained in Dulbecco's Modified Eagle Medium (DMEM, 08456-65 for MCF7/Fucci(SA)2.5, 08459-35 for MDA-MB-231-D/Fucci(SA)-Luc2, Nacalai Tesque). The culture medium was supplemented with 10% fetal bovine serum (FBS, 26140-079, gibco) and 1% penicillin-streptomycin (P/S, P4333-100ML, Sigma-Aldrich). Prior to on-chip imaging and library preparation, 4.0 × 10^5 of MCF7/Fucci(SA)2.5, and 2.0 × 10^5 of MDA-MB-231-D/Fucci(SA)-Luc2 cells were suspended in 200 μL Gene Pulser® Electroporation Buffer with 100 nM DNA tags and performed electroporation using Gene Pulser MxcellTM Electroporation System (Bio-Rad) with the following parameters: voltage of 200 V, capacitance of 2000 μF, resistance of ∞ Ohm, and duration of 20 ms for MCF7/Fucci(SA)2.5, and MDA-MB-231-D/Fucci(SA)-Luc2. Subsequently, we added 1 mL medium and cultured overnight in 24-well plate dishes, each containing cells with one of 16 different types of DNA tags. We treated the cells with palbociclib (S4482, Selleck Chemicals LLC) at various concentrations with 0.1% dimethyl sulfoxide for 24 h. After washing with 1× phosphate-buffered saline (PBS, 14249-24, Nacalaitesque), the cells were stained individually in each well with 16 different combinations of four CellTrace (2.5 μM Violet, 1 μM CFSE, 2.5 μM Yellow, and 0.2 μM Far Red) according to the manufacturer's protocol. We then suspended the 16 sub-pools of cells in a tube of imaging medium (DMEM, 08490-05, Nacalai Tesque, without FBS and P/S) at equal concentrations.
isolate
Mixture of mRNA from the MCF7/Fucci(SA)2.5 and MDA-MB-231-D/Fucci(SA)-Luc2 cells
age
not applicable
biomaterial_provider
MCF7/Fucci(SA)2.5: RIKEN BRC, MDA-MB-231-D/Fucci(SA)-Luc2: provided by Prof. Takeshi Imamura of Ehime University
collection_date
not applicable
geo_loc_name
not applicable
sex
not applicable
tissue
Breast; Mammary gland
cell_line
MCF7/Fucci(SA)2.5, MDA-MB-231-D/Fucci(SA)-Luc2
cell_type
epithelial cell
disease
Adenocarcinoma
sample_type
RNA and DNA tag
treatment
Palbociclib treatment at eight different concentrations ranging from 0 to 2000 nM for 24-h
BioSampleModel
Human