Notes: A novel target of non-steroidal anti-inflammatory drugs (NSAIDs) was identified using the Caspase-Glo® -1, Caspase-Glo® 3/7, and Caspase-Glo® 9 assays. The authors observed that an inhibition of caspase activity lead to decreased cell death and inflammation. The CellTiter-Fluor™ and CellTiter-Glo®  assays were using to monitor cell viability. (5163)

Notes: CYP450 activity was measured using the P450-Glo™ Assays for CYP3A4, CYP2B6 and CYP1A2, and viability was measured using two separate luminescent assays, one of which being CellTiter-Glo® Cell Viability Assay. (4877)

Notes: These authors investigated how autophagy controls haematopoietic stem cell (HSC) function, and how changes in autophagy levels affect HSC ageing. They showed that loss of autophagy in HSCs causes accumulation of mitochondria and an activated metabolic state. They used the Caspase-Glo® and CellTiter-Glo Assays to detect apoptosis and ATP, respectively, and used the Glucose Uptake-Glo™ Assay to study metabolic changes. (5018)

Notes: The number of viable cells in a culture was determined using the CellTiter-Glo® Luminescent Cell Viability Assay  (4874)

Notes: These authors report that glutamine promotes proliferation of cancer cells independently of its use as a metabolic fuel or as a precursor of glutathione, and that glutamine also activates transcription factors hypoxia-inducible factor-1, mammalian target of rapamycin and c-Myc, but these factors do not mediate the effects of glutamine on cancer cell proliferation. During the study, total RNA was isolated from cells using the Maxwell® RSC instrument,  ATP levels were measured using the CellTiter-Glo® Assay and intracellular glutamine and glutamate were measured using the Glutamine/Glutamate-Glo™ Assay. HIF-1 and PHD activities were measured using the Dual-Luciferase® Reporter Assay.

Notes: Viability was assessed using the CellTiter-Glo® Luminescent Cell Viability Assay. (4875)

Notes: Cell viability of IDH1 wild-type or mutant cell lines under treatment conditions was assessed using the CellTiter-Glo® Luminescent Cell Viability Assay. From these numbers, the IC₅₀ values were calculated. To evaluate apoptosis, 7,000–8,000 cells were treated with DMSO or compounds singly or in combination (12.5nmol/L NAMPT inhibitors and/or 200μmol/L temozolomide or 50μmol/L Z-VAD-FMK) for 6 or 24 hours and then measured using Caspase-Glo® 3/7 Assay. Changes in NAD+, NADH, NADP+, and NADPH were assessed using the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays. (4946)

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: In this paper, the RealTime-Glo™ and CellTiter-Glo® cell viability assays were used to measure viability of various breast cancer cell lines cultured in the OrganoPlate® microfluidic chip system from Mimetas. The toxicity of different chemicals on three different breast cancer cell types grown in 2D and 3D culture was evaluated. The RealTime-Glo™ assay was useful for measuring viability on day 0 prior to drug exposure and after 3 days of treatment. The CellTiter-Glo® Assay also was used, limited to the endpoint approach. (4934)

Notes: Relative levels of ATP were measured using CellTiter-Glo® Luminescent Cell Viability Assay. (4876)

Notes: 2D and 3D proliferation assays were performed using the CellTiter-Glo® Reagent with a GloMax® Discover System. (4745)

Notes: The rAAV-mediated genome editing technology was used to introduce a NanoLuc^® reporter sequence downstream of and in frame with the last coding exon of the HIF1A gene in HCT116 cells. This created a reporter cell line with a single allele of HIF1A endogenously tagged with a NanoLuc^® reporter fusion. The cell line was used to study compound inhibition of Hif1a signaling. NanoLuc^® luciferase activity was measured using Nano-Glo^® reagent. For qHTS, cells at 1500 cells/well in 1536-well plates were incubated with test compounds at 37°C, 5% CO₂, 1% O₂ for 18 hours in a humidified CO₂ incubator with variable oxygen control, followed by addition of Nano-Glo^® reagent or CellTiter-Glo^® cell viability assay reagent. The HRE-bla assay was conducted using the CellTiter-Glo^® reagent. HCT116 and ME-180 cell proliferation was quantified as relative luminescence unit (RLU) values using the CellTiter-Glo^® viability assay reagent. (4761)

Notes: The authors utilized phosphoproteomics to identify host factors that are activated upon Influenza A infection. In total, 1116 proteins showed changes in regulation including cyclin-dependent kinases. The authors tested the effects of cyclin-dependent kinase inhibitors on cell viability and caspase activation using the CellTiter Glo® and Caspase-Glo® assays. (5154)

Notes: Proliferation assays were performed using the CellTiter-Glo^® Luminescent Cell Viability Assay and the GloMax^®-Multi+ Microplate Reader. (5083)

Notes: The authors used the CellTiter-Glo® Cell Viability Assay and CellTox™ Green Cytotoxicity Assay to monitor the reproducibility of their acoustic dispensing. The authors report that adding CellTox™ Green Cytotoxicity Assay at plating "gives more solid signals than adding at the end of the assay” and speculate that the dye stabilizes the DNA of dead cells. The CellTiter-Glo® Assay data supports that the dye is nontoxic in the 72-hour incubation. The authors routinely predispense 12.5nl of CellTox™ Green with preplated compounds into 1536-well assay plates using an Echo 550 device and store plates under nitrogen gas in storage pods to produce assay-ready plates awaiting cell additions. The authors also report preliminary results of using the nonlytic RealTime-Glo® Viability Assay and find highly correlated results with the lytic CellTiter-Glo® Assay. The authors relate that use of the nonlytic RealTime-Glo® and CellTox™ Green Assays open the possibility of doing further work with the same cells. (4703)

Notes: This study investigated the reason why cancers become resistant to anti-cancer treatments over time. Non-small-cell lung cancers were cultured for weeks in the presence of EGF receptor inhibitors and resistant clones obtained based on a specific mutation in the EGF receptor. Clones that became resistant within 6 weeks were found to result from cells already containing the EGFR mutation. Clones taking about 24 weeks to become resistant evolved from the EGFR mutation. RealTime-Glo™ MT Cell Viability Assay was employed in several studies over a 10-week period to monitor proliferation (data presented in Supplementary Figures). Each week prior to media change, RealTime-Glo™ Assay reagents were added, incubated for 1 hour and luminescence measured. After 72-hour incubations, short-term viability was measured with the CellTiter-Glo® Luminescent Cell Viability Assay. (4699)

Notes: Mouse embryonic fibroblasts were treated with vehicle or DPQ for 1 hour followed by exposure to H₂O₂.  Intracellular NAD+ was measured with the NAD/NADH-Glo™ Assay and cellular ATP was measured with the CellTiter-Glo® Viability Assay. (4845)

Notes: The authors purified ctDNA from 1ml plasma using the Maxwell® RSC ccfDNA Plasma Kit with the Maxwell® RSC Instrument. Cell line mismatch repair deficiency was confirmed by the MSI Analysis System, Version 1.2. Cell line authentication was performed using the Cell ID™ and GenePrint® 10 Systems. DNA from cultured and treated cells were purified with the Wizard® SV and SV 96 Genomic DNA Purification Systems. Cetuximab-treated cells were measured for viability with the CellTiter-Glo® Luminescent Cell Viability Assay. (4777)

Notes: In this study, researchers used high-throughput compatible 384-well and 1536-well assay plate formats with spheroids of various cell types. Viability and apoptosis of monolayer and 3D tumor models was assessed. (5223)

Notes: The authors used bioluminescence resonance energy transfer (BRET) to test the ability of a bromodomain 9 ligand to disrupt binding to histone. HEK 293 cells were cotransfected with a histone H3.3-HaloTag® fusion vector and either NanoLuc®-BRD9 bromodomain or NanoLuc®-full-length BRD4 fusion vector. After 24 hours, the transfected cells were trypsinized, diluted in phenol red-free DMEM with or without 10nM of HaloTag® NanoBRET™ 618 Ligand and dispensed into a 96-well plate. One of two potential BRD-disrupting compounds, 7d or 11, was adding to a final concentration of 0.005–33μM, cells were incubated for 18 hours and NanoBRET™ Nano-Glo® Substrate (final concentration 10µM) was added. Fluorescence was measured and a corrected BRET ratio calculated. Cytotoxicity was assessed after the NanoBRET™ assay by incubating the cells with the CellTiter-Glo® Reagent for 30 minutes and measuring luminescence. To examine histone H3.3 localization, HEK 293 cells were transfected with the histone H3.3-HaloTag® fusion vector using FuGENE® HD Transfection Reagent. After 24 hours, cells were labeled with 5μM HaloTag® TMR ligand for 15 minutes before washing with complete medium, incubated for 30 minutes and imaged with a confocal microscope. (4568)

Notes: The authors describe a homogeneous, nonlytic, bioluminescent assay that measures cell viability in real time. They monitored cell health for 72 hours from the same test samples, distinguished differential cell growth, and investigated drug mechanism of action by analyzing time- and dose-dependent drug effects. The real-time measurements enabled them to detect cell death immediately (>75% signal decrease within 15 minutes of digitonin addition), analyze drug potency versus efficacy, and identify cytostatic versus toxic drug effects.They then screened an oncology compound library (Z′ = 0.7) and identified compounds with varying activity at different time points. (4590)

Notes: A549 and HCT116 cells were transfected with an inducible shRNA to reduce eEF2K levels. Culturing of the cells at various medium pH lead to a decrease in viability and increase in cytotoxicity, which was increased when the shRNA was induced. Cell viability was measured with the CellTiter-Glo® Luminescent Cell Viability Assay, and cytotoxicity was measured with the CellTox™ Green Cytotoxicity Assay. (4712)

Notes: The authors of this study use large 3D multipcellular spheroids (MCSs, >200µm) to serve as intermediate models between 2D cell culture and in vitro systems for investigating drug delivery and photodynamic therapy (PDT) for cancer. They evaluate four metal complexes that target the mitocondrion for PDT in 3D MCSc formed from HeLa cells. Cell viability is evaluated in the standard 2D culture system using the CellTiter-Glo® Cell Viability Assay, and in the 3D MCS using the CellTiter-Glo® 3D Cell Viability Assay. (4583)

Notes: Humans are exposed to a cocktail of low-dose chemicals in the environment, through diet, and through medication. However, most studies looking at compound toxicity, look at these compounds in isolation, not as they might be encountered in the environment—in mixtures. The pregnane X receptor (PXR) is a xenoreceptor that has been identified by the US Environmental Protection Agency as a major target of environmental and dietary chemicals, and many studies have highlighted the role of nuclear receptors like PXR in transducing the deleterious effects of endocrine disrupting compounds in the environment. The authors of this study used compound screening and functional analysis to demonstrate that the combined use of an environmentally persistent organochlorine pesticide and the active component of contraceptive pills (17α-ethinylestradiol) produces synergistic effects on PXR and expression of its target gene, the cytochrome P450 gene, CYP34A. Gene expression of CYP3A4 was measured using a luciferase reporter created in a pGL3-basic backbone. Activity of the CYP3A4 protein was measured in primary human hepatocytes using the P450-Glo™ CYP3A4 Assay with Luciferin-IPA, and cell number was normalized using the CellTiter-Glo® Luminescent Cell Viability Assay. (4579)

Notes: To avoid the trial-and-error methodology involved in developing multidrug treatments, the authors designed and tested a platform for high-throughput combination screening to easily discover pairs of small molecules that could then be explored further. Cell lines tested were dispensed at 1,000 cells per well in 5µl of medium into a 1,536-well plate either directly to compound matrix plates or adding 23nl of bortezomib and library compounds to the cells. After a 48-hour incubation, 3µl of CellTiter-Glo® Luminescent Cell Viability Assay reagent was dispensed and luminescence read to assess cell proliferation. (4209)