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Current Chemical Genomics 3, 33-41. In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high-throughput screening 2009

Niles, A.L., Moravec, R.A. and Riss, T.L.

Notes: The authors review the use of in vitro cytotoxicity testing in drug discovery to characterize the toxic potential of new chemical entities (nce) at the earliest stages of profiling. DOI: 10.2174/1875397300903010033 (4000)

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Toxicology in Vitro 23, 1170-1171. Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds 2009

Wu, Y., Connors, D., Barber, L., Jayachandra, S., Hanumegowda, U.M. and Adams, S.P.

Notes: The authors describe a multiplexed in vitro assay to detect nephrotoxicity and gain information about mechanism of cell death in HK-2 (human kidney-2) cells. The multiplexed assay involved an LDH assay to detect necrosis, a caspase-3/7 assay to detect apoptosis, a reazurin assay to assess metabolic state, and a DNA dye staining assay to monitor nuclear morphology. (4002)

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Mol. Cell. Endocrinol. 264, 50-60. Novel estrogen receptor beta transcript variants identified in human breast cancer cells affect cell growth and apoptosis of COS-1 cells. 2007

Treeck, O., Pfeiler ,G., Horn, F., Federhofer, B., Houlihan, H., Vollmer, A., and Ortmann, O.

Notes: This study identified two novel transcript variants of the estrogen receptor ERβ that were expressed in the ERα-negative breast cancer cell line MDA-MD-231. These variants were identified after amplification of ERβ transcripts from the breast cancer cell line by RT-PCR. The amplification products were then excised from gels and subcloned into the pTARGET™ Mammalian Expression Vector prior to sequencing. COS1 cells, which do not express the estrogen receptor, were then stably transfected with full-length ERβ or one of the splice variants and the effects on cell proliferation, apoptosis, and estrogen response were evaluated. In COS1 cells expressing either ERβ or the transcript variants cell proliferation decreased and basal apoptosis (caspase 3/7 activity) increased, compared to cells transfected with vector alone. Exposure to therapeutic doses of tamoxifen induced apoptosis in cells expressing the full-length ERβ but not in cells expressing either of the variant isoforms. (3618)

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FEBS J. 273, 2206–2222. Differing molecular mechanisms appear to underlie early toxicity of prefibrillar HypF-N aggregates to different cell types. 2006

Cecchi, C., Pensalfini, A., Baglioni, S., Fiorillo, C., Caporale, R., Formigli, L., Liguri, G., and Stefani, M.

Notes: The CellTiter-Blue® Cell Viability Assay was used to monitor viability of Hend murine endothelial cells and IMR90 fibroblasts in the presence of various concentrations (0.02, 0.2, 2.0 or 20 µm) of the N-terminal domain of the prokaryotic hydrogenase maturation factor (HypF-N). The cells were exposed to HypF-N for up to 24 hours before the cells were washed and cultured in fresh media. The CellTiter-Blue® Reagent was added and the cultures incubated for 1 hour before fluorescence readings were taken. Data was expressed as percent viability compared to a control. (3476)

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J. Biol. Chem. 281, 324-333. Mitochondrial transcription factor A induction by redox activation of nuclear respiratory factor 1. 2006

Piantadosi C.A., and Suliman, H.B.

Notes: Cell proliferation and viability of H4IIE rat hepatoma cells or HCC1937 human mammary primal ductal gland carcinoma cells were assessed with the CellTiter-Blue® Cell Viability Assay. Both cell lines were also exposed to 15µM of the lipophilic oxidant, tertiary butyl hydroperoxide (t-BOOH) for 5 hours. Incubations with the CellTiter-Blue® Cell Viability dye were carried out for 4 hours. Data are expressed as percent viability compared to control cultures. (3478)

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Nature 434, 652-658. Cyclophilin D-dependent mitochondrial permeability transition regulates some necrotic but not apoptotic cell death. 2005

Nakagawa, T., Shimizu, S., Watanabe, T., Yamaguchi, O., Otsu, K., Yamagata, H., Inohara, H., Kubo, T., Tsujimoto, Y.

Notes: In this study, the role of Cyclophilin-D (CypD) in the mitochondrial permeability transition (mPT) response was investigated using CypD-deficient mice. The CellTiter™ Blue Cell Viability Assay was used to measure the viability of mouse embryonic fibroblasts (MEF) and hepatocytes isolated from normal and CypD-deficient mice after exposure to various apoptotic stimuli and H2O2. (3398)

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Hum. Mol. Genet. 13(20), 2409-20. Correction of aberrant FGFR1 alternative RNA splicing through targeting of intronic regulatory elements 2004

Bruno, I.G., Jin, W., Cote, G.J.

Notes: Human U251 glioblastoma cell lines treated with antisense morpholino oligonucleotides were assessed for viability and apoptosis by multiplexing the CellTiter-Blue® Cell Viability and Apo-ONE® Homogeneous Caspase-3/7 Assays on single cell cultures. Cell viability was measured 4 hours after the addition of the CellTiter-Blue® Cell Viability Reagent to the cultures. Next, apoptosis measurements were performed on the same cell cultures by adding Apo-ONE® Homogeneous Caspase-3/7 Assay reagent to the cultures. Caspase-3/7 activity was then measured 12 hours later. (3168)

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Assay Drug Dev. Technol. 3, 469-477. HTS Compatible Assay for Antioxidative Agents Using Primary Cultured Hepatocytes 2003

Gaunitz, F. and Heise, K.

Notes: Primary hepatocytes from Sprauge-Dawley rats were incubated with various concentrations of tert-butylhydroperoxide to induce oxidative stress and then cell viability, cytotoxicity and apoptosis were assayed with Promega's CellTiter 96 AQueous One Solution Cell Proliferation Assay, CellTiter-Blue Cell Viability Assay, CellTiter-Glo Luminescent Cell Viability Assay, CytoTox-ONE Homogeneous Membrane Integrity Assay, Apo-ONE Homogeneous Caspase-3/7 Assay and assay systems. (2969)

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