Notes: The NADPH/NADP and GSH/GSSG ratios were measured in cells to determine that CPT1A is required to maintain ROS homeostasis. (5039)

Notes: The authors examine the effect of co-treatment with dtBHQ and sulforaphane on expression of the transcription factor Nrf2 and regulation of antioxidant genes. The GSH/GSSG-Glo™ assay kit was used to measure levels of oxidative stress in HaCaT cells after dtBHQ treatment. Activation of antioxidant response element genes was measured with the Dual-Luciferase^® Reporter Assay. Interestingly, sulforaphane treatment in the presence of reactive oxygen species leads to activation of antioxidant genes while inhibiting Nrf2 protein expression. (5176)

Notes: The CellTiter-Glo^® assay was used in a high-throughput drug screening assay to identify compounds which increase mitochondrial ATP production in muscle cells. Hesperetin was found to both increase mitochondrial function and reduce oxidative stress as measured by the GSH/GSSG-Glo™ assay. The authors present a novel screening platform to identify possible therapeutics for sarcopenia and other mitochondrial dysfunction diseases. (5177)

Notes: The authors use whole transcriptome analysis to investigate monocyte expression profiles in X-linked adrenoleukodystrophy. Viability and glutathione oxidative balance of monocytes was measured using the MultiTox-Glo and GSH/GSSG-Glo™ assays on the GloMax^® system. (5178)

Notes: The Nrf2-response element luciferase was used to measure transcriptional activity of a novel transcriptional activator Plant Homeodomain Finger 2 (Phf2). Interestingly, Phf2 was identified to protect the liver from Non Alcoholic Fatty Liver Disease (NAFLD) through histone demethylation. CGSH/GSSG-Glo™ was used to monitor glutathione content in the liver, while the ApoTox-Glo™ assay was used to monitor for cell viability and toxicity. (5179)

Notes: In this study, the CellTiter-Glo® 2.0 Assay was used to determine viability after treatment of cells with various compounds, and the GSH/GSSG-Glo™ Assay used to measure glutathione levels. The pGL3 Vectors and Dual-Luciferase® Assay were used in Reporter Assays. (5006)

Notes: B16 F10 murine melanoma cells were treated with fenofibrate and the ratios of NADP+/NADPH and GSH/GSSG were examined after PPARa knockdown or over-expression. The NADP+/NADPH levels did not change appreciably but the GSH/GSSG levels did fluctuate. The study used the NADP/NADPH-Glo™ and GSH/GSSG-Glo™ Assays. Assays were read on a GloMax® 20/20 Luminometer. (4852)

Notes: The levels of NADP+ and NADPH in dorsal root ganglion neurons were followed as an indicator of the pentose phosphate branch of glycolysis. Meclizine helped preserve the NADP+/NADPH ratio in hypoxia but was unable to preserve the ratio in the presence of 2-deoxyglucose. As an indicator of the utilization of NADPH, the GSH/GSSG ratio was monitored in the cells.  Meclizine induced increased GSH production during hypoxia which of course was abrogated by 2-deoxyglucose. The study used the NADP/NADPH-Glo™ and GSH/GSSG-Glo™ Assays. (4849)

Notes: The effect of PHGDH knockdown on redox homeostasis in human breast cancer cell lines was examined with regard to GSH/GSSG ratios and NADPH levels under normoxic and hypoxic conditions. The study used the NADP/NADPH-Glo™ and GSH/GSSG-Glo™ Assays. (4850)

Notes: A triple drug treatment-parthenolide, 2-deoxyglucose, and temsirolimus (PDT); each chosen for the ability to inhibit the pentose phosphate pathway and the Nrf-2-mediated anti-oxidant reponse-was tested against AML cells, leukemic stem/progenitor cells and normal heamtopoietic counterparts. The PDT treatment was potently toxic to the AML and leukemic stem/progentor cells with little toxicity to the normal cells. The NADPH and GSH/GSSG ratios were monitored in this study with the NADP/NADPH-Glo™ Assay and GSH/GSSG-Glo™ Assay.  (4856)

Notes: The authors of this study sought to determine whether cucurbitacin B has antitumor activity against the ovarian cancer cell line A2870 and whether it can sensitize the cisplatin-resistant cell line A2780CP to treatment. They compared caspase activity in A2780CP cells either preincubated with cucurbitacin B before treatment with cisplatin or cells not pretreated using the Caspase-Glo® 3/7 Caspase Assay. Oxidized and total glutathione were measured from both cell lines (before and after cisplatin exposure, with and without preincubation with cucurbitacin B) using the GSH/GSSH-Glo™ Glo Assay. The level of reactive oxygen species in cell lines was also measured by detecting ROS converted to H₂O₂ using the ROS-Glo™ Assay. Cell Viabilty of 3D spheroids formed from the A2780CP cell line was assessed using the CellTiter-Glo® 3D Cell Viability Assay. (4581)

Notes: The role of RBM45 in the KEAP1/NRF2/ARE pathway was examined through transfection of HEK293 cells with either overexpression vectors or siRNAs. Overexpression of RBM45 increased ROS levels in cells with a concurrent increase in GSSG levels in comparison to GSH levels with a concurrent increase in cytotoxicity. Overexpression of RBM45 with a deleted nuclear localization signal slightly decreased the GSSG levels. The cytotoxic effect was abrogated with an siRNA directed to RBM45. GSH and GSSG levels were measured with the GSH/GSSG-Glo™ Assay, and cytotoxicity was measured with the CellTox™ Green Cytotoxicity Assay using the end-point protocol. (4713)

Notes: 5-aminolevulinic acid (5-ALA)-induced protoprophyrin IX (PpIX) fluorescence is often used for diagnostics and therapy in malignant glioma surgery; however changes in the ability of malignant glioma cells to accumulate PpIX can result in erroneous interpretation of the fluorescent signal and affect the safety of fluorescence-guided surgery. Such changes can result from several factors, including drug interactions. The authors of this paper investigated the effects of antiepiletic drugs (AED) commonly given to glioma patients on the ability of glioma cells to accumulate PpIX. They looked at cell health and metabolism in glioma cells treated with these drugs, including glutathione metabolism. The GSH/GSSG-Glo™ Assay was used to generate a ratio of reduced (GSH) and oxidized (GSSG) glutathione in glioma cell lines (U-87 MG and U373 MG). Cells were seeded at 5 × 10³ cells/well in 96-well plates and treated with either 47µg/ml phenytoin, 104µg/ml levetiracetam or DMSO-only for three days, and the assay was performed as described in the technical manual. Neither test compound effected the GSH/GSSG ratio.  (4211)