MCF7 and T47D cells with a doxycycline (DOX)-inducible Y537S or D538G (DOX-Y537S or DOX-D538G) mutation in ER (6 (link)), were grown in DMEM and RPMI, respectively [supplemented with 10% FBS, 10 μg/mL penicillin-streptomycin-glutamine (PSG), and 500 μg/mL Geneticin], hereafter referred to as full media (FM) conditions. DOX-inducible ER-mutant MCF7 and T47D cells (MCF7 DOX-Y537S, MCF7 DOX-D538G, and T47D DOX-Y537S) were treated with 1 μg/mL DOX in FM for 3 days to induce the expression of the mutation and subsequently maintained in phenol-red-free DMEM or RPMI supplemented with 10% charcoal-stripped FBS, PSG, and Geneticin, hereafter referred to as HD conditions. MCF7 cells with a stable TALEN knock-in Y537S (KI-Y537S) or D538G (KI-D538G) ER mutation (6 (link)) were grown in DMEM supplemented with 10% FBS and PSG. Palbociclib-sensitive (PalboS) and palbociclib-resistant (PalboR) T47D cells (23 (link)) and MCF7 cells (24 (link)) were grown in DMEM supplemented with 10% FBS and PSG, with or without palbociclib 100 nmol/L for the PalboS cells, and with palbociclib 1 μmol/L for the PalboR cells. Cells were authenticated by short tandem repeat profiling (Bio-Synthesis) and regularly tested for Mycoplasma contamination using the MycoAlert Mycoplasma Detection Kit (Lonza) according to the manufacturer's instructions.
Short tandem repeat profiling
Manufactured by Bio-Synthesis
Short tandem repeat profiling is a genetic analysis technique used to examine specific regions of the human genome. It involves the detection and analysis of short, repeated DNA sequences, which can provide valuable information for various applications, such as forensics, paternity testing, and population genetics studies.
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Lab products found in correlation
2 protocols using short tandem repeat profiling
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Engineered ER-mutant Breast Cancer Cell Lines
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Cell Authentication and Compound Sourcing
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The cell lines used in this study and their sources are listed in Supplementary Table S1. Cell lines were authenticated by short tandem repeat profiling (Bio-Synthesis Inc.) or were used within 5 passages of original purchased stocks. The cells were confirmed to be free of mycoplasma by PCR (CLEA Japan, Inc.). AZD4547 was synthesized at Taiho Pharmaceutical Co., Ltd., infigratinib and pemigatinib were purchased from MedChemexpress Co., Ltd., and erdafitinib was purchased from Namiki Shoji Co., Ltd.
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