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2 protocols using doxcycline

1

Inducible Wild-type p53 Expression in Triple Negative Breast Cancer Cells

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The human breast cancer cell line HCC1937 was purchased from American Type Culture Collection (ATCC). The HCC1937 cells were negative for expression of ER, PR, and HER2, referred to as a triple negative tumor, and have mutations of TP53 and BRCA1. HCC1937 cells were stably transfected with a wt-p53-inducible plasmid (Tet-on Advanced system, Clontech, USA), and one of the isolated clones was designated as HCC1937/p53 and used for the experiments. The HCC1937/p53 cells were cultured in RPMI1640 (Nacalai tesque, Kyoto, Japan), containing 10% FBS (SIGMA, USA), and ZeocinTM (1 μg/mL, InvivoGen, USA). The HCC1937/p53 cells were cultured on APS-coated slides (MATSUNAMI, Japan) in doxycycline (Takara, 1 ng/mL)—containing media for 1–10 days, and those cells treated with doxcycline for 1 day were designated as dox1d.
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2

Establishing Triple-Negative Breast Cancer Model

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The human breast cancer cell line HCC1937 was purchased from American Type Culture Collection (ATCC). The HCC1937 cells were negative for expressions of ER, PR, and HER2, referred to as a triple-negative tumor, and had mutations of TP53 and BRCA1 [24 (link)]. HCC1937 cells were stably transfected with a wt-p53-inducible plasmid (Tet-on Advanced System, Clontech, USA), and one of the isolated clones was designated as HCC1937/p53 and used for the experiments. The HCC1937/p53 cells were cultured in RPMI1640 (Nacalai Tesque, Kyoto, Japan), containing 10% fetal bovine serum (FBS) (SIGMA, USA) [18 (link)], and ZeocinTM (1 μg/mL, InvivoGen, USA). The HCC1937/p53 cells were cultured in doxycycline (Takara, 1 ng/mL)-containing media for 1–7 days, and those cells treated with doxcycline for 2 days were designated as Dox2d [20 (link)].
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