Notes: These researchers demonstrated that thyroid hormones induce CYP3A4 metabolic activity, and this modulation may regulate the metabolism of a chemotherapeutic agent. (5211)

Notes: Hypoxic cancer cells constitute a major challenge in chemo- and radiotherapy. The researchers present evaluation of APD-CLD compounds that induce selective cell death in hypoxic cancer cells by impeding mitochondrial function. The RealTime-Glo™ Annexin V Apoptosis Assay, CellTiter-Blue® Cell Viability Assay, CellTox™ Green Real-Time Cytotoxicity Assay and GSH-Glo™ Glutathione Assay were used to assess mechanism of action. (5168)

Notes: This study investigated the effect of the Bacillus thuringiensi toxins parasporin 3 on HepG2 cells. Understanding how Bt toxins target human cell lines has implications for the risk assessment of products containing these toxins, such as transgenic, insect-resistant plants. The authors used the CellTiter®-Blue, CellTiter®-Glo, Caspase-Glo® and ROS-Glo™ assays to assess viability, apoptosis induction, and oxidative stress in cultured cells exposed to the toxin. (5038)

Notes: SR-786 NPM/ALK-positive human anaplastic large cell lymphoma cell line was treated with lobatin B, a plant-derived natural compound. Total RNA was isolated with the ReliaPrep™ RNA Cell Miniprep System and reversed transcribed into cDNA with the GoScript™ Reverse Transcription System. Levels of NPM/ALK and NF-κB expression was measured with the dye-based GoTaq® qPCR System. SR-786 cells were also analyzed for functioning metabolism with the CellTiter-Blue® Cell Viability Assay and apoptosis with the Apo-ONE™ Homogeneous Caspase-3/7 Assay. (4604)

Notes: A phenotypic screen of a 25,000 compound library for inhibitors of U2OS osteosarcoma cell line growth identified two compounds that showed activity of osteosarcomas and not HepG2 or primary human osteoblasts.  Cell Viability in the primary and secondary screens was monitored with the CellTiter-Blue® Cell Viability Assay.  Subsequent studies identified the mechanism of cell death as apoptosis as judged by the Caspase-Glo® 3/7 Assay and cytotoxicity measured with the CellTox™ Green Cytotoxicity Assay.
(4649)

Notes: Primary normal human bronchial epithelial (NHBE) cells were cultivated on semi-permeable membranes of cell culture inserts exposed directly to vapor from 200 puffs of two different e-cigarette liquids (0% and 2.4% nicotine). Twenty-four hours post exposure, two assays were multiplexed to assess oxidative stress (ROS-Glo™ H₂O₂ Assay) and cell viability (CellTiter-Blue® Cell Viability Assay). First, the cells were incubated with 200µl of medium + 50µl of H₂O₂ Substrate Solution for 3 hours and 75µl of the mixture transferred to a white 96-well plate. An equal volume of Detection Solution was added, incubated for 20 minutes at room temperature and luminescence detected. The exposed cells had the remaining medium + H₂O₂ Substrate Solution removed and replaced with 300µl of fresh medium + 60µl of CellTiter-Blue® Reagent. After a 2-hour incubation, 100µl of solution was transferred to a black 96-well plate and fluorescence measured at 544nm_Ex/590nm_Em. (4569)

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)

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)

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)

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)

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)

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)

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)

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)