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Hum. Mol. Genet. 21(3), 664-680. An aggregation sensing reporter identifies leflunomide and teriflunomide as polyglutamine aggregate inhibitors. 2012

Fuentealba, R.A., Marasa, J., Diamond, M.I., Piwnica-Worms, D., and Weihl, C.C.

Notes: These authors developed a protein-aggregation reporter that uses huntingtin exon 1 containing 72 glutamines fused to the N-terminal end of firefly luciferase (httQ72-Luc). This construct did not aggregate unless seeded by a non-luciferase-containing polyglutamine (polyQ) protein. Upon co-aggregation, httQ72-luc becomes insoluble and loses its enzymatic activity.  httQ72-Luc and a polyQ protein were used to screen a library for compounds that prevent polyQ aggregation. Leflunomide and its active metabolite teriflunomide inhibited protein aggregation independently of their known role in pyrimidine biosynthesis. This study demonstrated the usefulness of luciferase-based protein aggregate reporters for high-throughput screening applications. Luciferase activity was measured using the Luciferase Assay System and the GloMax®-Multi Detection System. (4188)

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J. Biotechnol. 157(4), 598-604. Detection of toxic lignin hydrolysate-related compounds using an inaA::luxCDABE fusion strain. 2012

Lee, S., and Mitchell, R.J.

Notes: This study evaluated use of a luciferase reporter construct for detection of hydrolysate related phenolic compounds generated during fermentation of plant products. The authors used a transcriptional fusion of the inaA promoter with the luxCDABE operon to detect phenolic compounds generated in plant hydrolysates. The GloMax® Multi Detection System was used to measure bioluminescence in various bacterial cultures. (4194)

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Biochim. Biophys. Acta 1822, 248-260. MiR-21 is involved in cervical squamous cell tumorigenesis and regulates CCL20. 2012

Yao, T., and Lin, Z.

Notes: This study identified that miR-21 is significantly overexpressed in human cervical squamous cancer tissues and cell lines, and showed that the level of miR-21 correlates with tumor differentiation, and regulates proliferation, apoptosis, and migration of HPV16-positive cells. The authors used gene expression profiling and luciferase reporter assay to identify candidate target genes for miR-21. Wildtype and mutant 3′-UTR sequences from the target gene CCL20 containing putative binding sites for miR-21 were subcloned into the psiCHECK-2 Vector and used to transfect HEK293 cells. The Dual-Luciferase® Reporter Assay System and GloMax® Multi Luminometer were used to measure luciferase activity from both constructs. (4192)

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Food Chem. Toxicol. 50(2), 116-123. Reactive oxygen species and PI3K/Akt signaling play key roles in the induction of Nrf2-driven heme oxygenase-1 expression in sulforaphane-treated human mesothelioma MSTO-211H cells. 2012

Lee, Y.J., Jeong, H.Y., Kim, Y.B., Lee, Y.J., Won, S.Y., Shim, J.H., Cho, M.K., Nam, H.S., and Lee, S.H.

Notes: This study investigated the relationship between nuclear factor erythroid-derived 2 related factor 2 (Nrf2)/heme oxygenase (HO)-1 induction, cell viability, and suforaphane treatment in human mesothelioma MSTO-211H cells. The GloMax® Multi Detection System was used to measure cell viability along with a formazan-based cell viability assay. (4193)

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Cancer Res. 72, 810-820. SMYD3 Promotes Cancer Invasion by Epigenetic Upregulation of the Metalloproteinase MMP-9. 2012

Cock-Rada, A.M., Medjkane, S., Janski, N., Yousfi, N., Perichon, M., Chaussepied, M., Chluba, J., Langsley, G., and Weitzman, J.B.

Notes: These authors investigated the role of matrix metalloproteinase (MMP-9) in a reversible model of cancer that is initiated by infection with intracellular Theileria parasites. They found that gene induction by parasite infection was associated with trimethylation of histone H3K4 (H3K4me3) at the MMP-9 promoter. The H3K4 methyltransferase SMYD3 was the only histone methyltransferase upregulated upon infection. They therefore investigated the role of SMYD3 overexpression on MMP-9 expression and cell migration, identifying SMYD3 as an important new regulator of MMP-9 transcription. During the study they used the GloMax® Multi Luminometer to measure luminescence and absorbance in reporter and cell viability assays. They also used the Dual-Luciferase® Reporter Assay to measure SMYD3 activity in cells transfected with a SMYD3 reporter, and the pGL4-hRluc/TK plasmid for normalization of the experimental reporter activity. GoTaq® DNA polymerase was used in semi-quantitative RT-PCR. (4189)

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African J. Agri. Res. 6, 3768–74. Effects of selective medium on lipid accumulation of chlorellas and screening of high lipid mutants through ultraviolet mutagenesis. 2011

Deng, X., Li, Y and Fei, X.

Notes: Three species of Chlorella microalgae of interest for use in renewable biofuels were cultured in four selective media, and the effects on growth and lipid content measured. To determine the lipid content of microalgae, the cells were stained with a final concentration 0.1μg/ml of Nile Red for 10 minutes. The GloMax®-Multi Detection System measured fluorescence using 470nm (excitation) and 570nm (emission) wavelengths. (4232)

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J. Biomed. Biotechnol. 757960, epub. The Over-expression of the β2 Catalytic Subunit of the Proteasome Decreases Homologous Recombination and Impairs DNA Double-Strand Break Repair in Human Cells 2011

Collavoli, A., Comelli, L., Cervelli, T., and Galli, A.

Notes: Proteasome activity was determined in cells expressing the β2 proteasome subunit using the Proteasome-Glo™ Trypsin-Like Cell-Based Assay. The authors determined the proteasome activity in the cells after exposure to the proteasome inhibitor MG132. Luminescence indicative of proteasome activity was measured using a GloMax®-Multi Detection System Luminometer. (4195)

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PLos ONE 6(6), e20994. Antimetastatic Effects of Phyllanthus on Human Lung (A549) and Breast (MCF-7) Cancer Cell Lines. 2011

Lee S.H., Jaganath I.B., Wang S.M., and Sekaran, S.D.

Notes: These authors investigated the ability of Phyllanthus plant extracts to affect the metastatic activity of human lung (A549) and breast (MCF-7) cancer cell lines. They initially used CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay and the GloMax®-Multi Detection System absorbance module to determine cytotoxicity of various Phyllanthus extracts. After determining the effective dose, the authors investigated the ability of these plant compounds to inhibit/reduce metastatic activity. They then evaluated the mechanism of cell death in treated cells using the Caspase-Glo® 3/7 Assay and the Glomax® Multi Detection System luminescence module to measure caspase activity, and the CytoTox-ONE™ Homogeneous Membrane Integrity Assay to measure LDH activity. (4196)

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Nucl. Acids Res. 39(3), e16. Characterization of L1 retrotransposition with high-throughput dual-luciferase assays. 2011

Xie, Y., Rosser, J.M., Thompson, T.L., Boeke, J.D., and Wenfeng, A.

Notes: This paper describes a rapid dual-luciferase-based assay for L1 retrotransposition that is amenable to high-throughput screening. A firefly luciferase vector in which the luciferase gene was disrupted by an antisense intron was constructed by introducing a 900-bp fragment of the human γ-globin intron into pGL4.13. This Fluc gene, interrupted by an antisense intron, gives only minimal luciferase expression unless the luciferase gene is restored by a retrotransposition event. The authors also tested a similar retrotransposition reporter using the pGL4.73 Renilla luciferase vector, but found that the firefly construct gave much higher signals. They therefore used the firefly luciferase retrotransposition reporter, a Renilla luciferase normalization control and the Dual-Luciferase® Assay to characterize profiles of retrotransposition by various human and mouse L1 elements, and to measure the kinetics of L1 retrotransposition in cultured cells. The GloMax® Multi Luminometer was used to quantify luciferase activity. (4205)

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RNA 17(3), 419-28. Differential utilization of decapping enzymes in mammalian mRNA. 2011

Li, Y., Song, M., Kiledjian, M.

Notes: This study analysed the roles of Dcp2 and Nudt16 in nonsense-mediated mRNA decay miRNA-mediated silencing. The authors used various luciferase reporter constructs to evaluate the significance of Dcp2 and Nudt16 in miRNA- and siRNA-mediated gene silencing in wildtype and knockout MEF cells. Various Renilla luciferase constructs containing or lacking miRNA target sites were cotransfected along with a control plasmid encoding firefly luciferase (for normalization purposes). Renilla and firefly luciferase luminescence were measured using the Dual-Luciferase® Reporter Assay and the GloMax®-Multi Luminometer. (4204)

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Atherosclerosis 216(1), 44-53. Nitric oxide activates PI3-K and MAPK signalling pathways in human and rat vascular smooth muscle cells: influence of insulin resistance and oxidative stress. 2011

Doronzo, G., Viretto, M., Russo,, I., Mattiello, L., Di Martino, L., Cavalot, F., Anfossi, G., Trovati, M.

Notes: To evaluate whether vascular smooth muscle cells from obese rats exhibit increased oxidative stress in comparison with cells from lean rats, these authors used a lucigenin-enhanced chemiluminescence method based on light emission from a reaction between reduced lucigenin and superoxide anion. The GloMax®-Multi Detection System luminescence module was used to quantify luminescence output. (4208)

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African J. Biotech. 10, 11600–10. Photosynthetic efficiency and lipid accumulation are affected by the concentration of carbon in microalgae Micractinium pusillum Y-002. 2011

Deng, X. et al.

Notes: A strain of microalgae, Micractinium pusillum strain Y-002, was grown in four different complete media as well as medium lacking one nutrient (nitrogen, iron, calcium, sulfur, phosphorus, magnesium, potassium or calcium). To determine the effect of the growth medium on lipid content of the microalgae, the cells were stained with a final concentration 0.1μg/ml of Nile Red for 10 minutes. The GloMax®-Multi Detection System measured fluorescence using 470nm excitation and 570nm emission wavelengths. (4233)

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African J. Microbio. Res. 5, 260-70. The effects of nutritional restriction on neutral lipid accumulation in Chlamydomonas and Chlorella. 2011

Deng, X., Fei, X. and Li, Y.

Notes: Microalgae strains Chlamydomonas reinhaditti CC124 and Chlorella vulgaris Y-019 were grown in four different media lacking elemental nutrients to examine the effect on oil production. To assay the lipid content of microalgae, the cells were stained with a final concentration 0.1μg/ml of Nile Red for 10 minutes. The GloMax®-Multi Detection System measured fluorescence using 470nm excitation and 570nm emission wavelengths. (4234)

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