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: Mid-exponential growth phase Lactococcus lactis cultures (OD600=0.6) were quenched in liquid nitrogen then stored at –20°C until assay. Extraction and quantification of NADH and NAD+ were performed with the NAD/NADH-Glo™ Assay. (4841)

Notes: The effect of KPT-9274 on RPTEC, Caki-1 and 786-0 cell NAD+/NADH levels was assessed with the NAD/NADH-Glo™ Assay. The treatment was performed on 3,000 cells per well for 48 hours prior to assay. (4840)

Notes: The effect of E4F1 on metabolism was assessed by comparing metabolites in primary fibroblasts from E4F1-knockout and control mice. The NAD+/NADH ratio was measured with the NAD/NADH-Glo™ Assay. (4842)

Notes: The effect of sirtuin2 overexpression on Leishmania infatum NAD+ homeostasis was assessed with the NAD/NADH-Glo™ Assay. (4844)

Notes: The NADP/NADPH-Glo™ Assay was used to look at variations in the level of total NADP+ and NADPH in primary porcine aortic endothelial cells with knockdown of von Willebrand Factor and the presence or absence of angiotensin II. The assay was read with a GloMax® Instrument. (4853)

Notes: The study found that cells undergoing IKKβ-knockdown had reduced cellular NADPH levels as judged through use of the NADP/NADPH-Glo™ Assay. Data are presented in the supplement. (4857)

Notes: The NAD/NADH-Glo™ Assay was used to monitor the NAD/NADH ratio of SKOv3 and SKOv3-ARHI cells with ARHI induction and glutaminase inhibition. (4836)

Notes: The NADP+/NADPH ratio was monitored in two cell lines, one with a glycolytic profile, MB-231, and an oxidative profile, SiHa, to monitor the effects of 6-AN treatment. The ratio was used as a measure of the pentose phosphate pathway. The ratio was monitored with the NADP/NADPH-Glo™ Assay. (4848)

Notes: One ml of 0.2 OD600 Mycobacterium tuberculosis (Mtb H37Rv) was treated with laterosporulin10 for 10 or 60 minutes then lysed with the assistance of a bead beater. The cleared lysate was analyzed for NAD/NADH and NADP/NADPH ratios using the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays. Both ratios fell dramatically upon treatment. (4827)

Notes: The effect of COUP-TFII on MPC1 promoter was evaluated with the Dual-Luciferase® Reporter Assay System. The effect of COUP-TFII on NADP+/NADPH levels in multiple prostate cancer cell lines was monitored with the NADP/NADPH-Glo™ Assay. The reduction in the ratio is likely due to the reduction in pentose phosphate shunt from lower glycolysis. (4847)

Notes: The NAD/NADH-Glo™ Assay was used to assess the effects of oxaloacetate, malate with or without glucose present on NAD+/NADH levels in SH-SY5Y cells at 20,000 cells per well. (4838)

Notes: Asparaginase treatment caused a significant increase in the NAD+/NADH ratio in all cell lines (data in supplement). The dinucleotide ratio was determined with the NAD/NADH-Glo™ Assay. (4839)

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 hRLuc gene from the pGL4.70[hRLuc] Vector was used as the source of Renilla luciferase used to make stably transfected Leishmania mexicana or donovani cell lines. The Renilla luciferase expression was used to monitor parasite loads  of infected cells. Luciferase activity was measured with the Renilla Luciferase Assay System. Amastigotes were treated with endochin-like quinolones and the ratio of NAD/NADH was measured with the NAD/NADH-Glo™ Assay.   (4837)

Notes: BEAS2B human bronchial epithelial cells were cultured for months in the presence of cigarette smoke condensate. The cells showed metabolic reprogramming including changes in the NADPH/NADP+ ratio. The measurement was made with the NADP/NADPH-Glo™ Assay and read on a GloMax® Instrument.  (4855)

Notes: The NADP/NADPH-Glo™ Assay was used to assess cellular utilization of the pentose phosphate pathway through monitoring NADPH levels. Use of a glucose-6-phosphate dehydrogenase inhibitor, 6-AN, effectively reduced NADPH levels. Data are shown in the supplemental material.  (4854)

Notes: The NAD/NADH-Glo™, NADP/NADPH-Glo™ and ENLITEN™ ATP Assays were used to monitor differences in ATP levels as well as NAD/NADH and NADP/NADPH ratios in primary  normal and cancerous colon cells in the various stages of the cell cycle. Each assay used 2 ×10⁵ cells per well and were normalized to total protein. (4826)

Notes: Mycobacterium tuberculosis grown as a biofilm or planktonic cells were found to have similar NAD/NADH and NADP/NADPH levels. The ratios were measured with the NAD/NADH-Glo™ and NADP/NADPH-Glo™ Assays. (4828)

Notes: Human megakaryocytes, with control or knockdown YAP, were analyzed for NAD+/NADH with the NAD/NADH-Glo™ Assay. (4843)

Notes: Serine/Glycine biosynthesis was examined for a panel of 79 non-small cell lung carcinomas, and each could be classified into a high-synthetic and a low-synthetic group based on expression of four enzymes involved in serine/glycine synthesis from glucose. ATF4 was identified as a key mediator of NRF2 activity for serine/glycine biosynthesis through transcriptional activation and not regulation of ATF4 translation (the latter being examined in a dual-luciferase reporter assay using Dual-Glo® Luciferase Assay System). A byproduct of the conversion of serine to glycine is production of NADPH. Seven high-synthetic cell lines exhibited a marked decrease in NADPH levels with regard to NADP+ level when the rate-limiting serine/glycine synthetic enzyme was knocked down with shRNAs with little change in NADPH/NADP+ ratios in six low-synthetic cell lines. The levels of NADPH and NADP+ were monitored with the NADP/NADPH-Glo™ Assay. Expression of the various shRNAs used in the study did not induce cytotoxicity as judged by staining with the CellTox™ Green Cytotoxicity Assay and visual examination by fluorescent microscopy. The identity of all cell lines used in this study were verified by STR analysis with a PowerPlex® System. (4715)

Notes: Induced pluripotent stem cells (iPSC) were differentiated into neurons and treated with 20, 100 and 500μM of ketamine for 6 and 24 hours to examine the neurotoxic effects of ketamine. The ApoTox-Glo™ Triplex Assay was used to examine cell viability and caspase-3/7 activation in the ketamine-treated iPSC-derived neurons. Fluorescence (cell viability) and luminescence (caspase-3/7 assay) was detected using a GloMax® multimode instrument. To examine if reactive oxygen species levels changed when iPSC-derived neurons were exposed to ketamine, the authors used the ROS-Glo™ H₂O₂ Assay with cells that were ketamine treated with or without a ROS scavenger for 6 and 24 hours. Luminescence was detected using a GloMax® instrument. Caspase activity was confirmed with or without the ROS scavenger using the Caspase-Glo® 3/7 Reagent from the ApoTox-Glo™ Triplex Assay. The levels of ketamine-induced oxidative stress were assessed in iPSC-derived neurons using the NAD/NADH-Glo™ Assay, and cellular ATP levels determined using the Mitochondrial ToxGlo™ Assay. The luminescence from both assays were measured on a GloMax® detection instrument. (4570)

Notes: Cell pellets of A375 human melanoma cells were lysed by passing through a needle, centrifuged to remove debris and protein quantitated. Twenty micrograms of protein from control and treated lysates were digested with 1µg of Sequencing Grade Modified Trypsin and used for mass spectrometry analysis. A375 cells (5 × 10⁵) were grown in a 10cm dish and treated for 24 hours before isolating RNA and DNA synthesized using M-MLV Reverse Transcriptase. The cDNA was then used in qPCR. A375 cells were plated at 3 × 10³ in 96-well half-volume white-wall plates, grown and treated for 24 hours. NAD, NADH and NADPH were determined using the NAD(P)H-Glo™ Detection System or NAD/NADH-Glo™ Assay and luminescence measured. (4963)