title: Development of a Multicolor Bioluminescence Imaging Platform to Simultaneously Investigate Transcription Factor NF-κB Signaling and Apoptosis.
abstract: Here we describe a novel multicolor bioluminescent imaging platform that enables us to simultaneously investigate transcription factor nuclear factor-κB (NF-κB) signalling and apoptosis. We genetically modified the human breast cancer cell line MDA-MB-231 to express green, red, and blue light-emitting luciferases to monitor cell number and viability, NF-κB promoter activity, and to enable specific cell sorting and detection, respectively. Z-DEVD-animoluciferin, the pro-luciferin substrate, was used to determine apoptotic caspase 3/7 activity. We used this multicolored cell line for the in vitro evaluation of natural compounds and in vivo optical imaging of tumor necrosis factor (TNFα)-induced NF-κB activation (Mezzanotte et al., PLoS One 9:e85550, 2014).
authors.affiliation: Department of Radiology, Leiden University Medical Center, Albinusdreef 2, PO Box 9600, 2300RC, Leiden, The Netherlands.
authors.forename: Vicky T
authors.initials: VT
authors.lastname: Knol-Blankevoort
authors.affiliation: Department of Radiology, Leiden University Medical Center, Albinusdreef 2, PO Box 9600, 2300RC, Leiden, The Netherlands.
authors.forename: Laura
authors.initials: L
authors.lastname: Mezzanotte
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Martijn J W E
authors.initials: MJ
authors.lastname: Rabelink
authors.affiliation: Department of Radiology, Leiden University Medical Center, Albinusdreef 2, PO Box 9600, 2300RC, Leiden, The Netherlands.
authors.forename: Clemens W G M
authors.initials: CW
authors.lastname: Löwik
authors.affiliation: Department of Radiology, Leiden University Medical Center, Albinusdreef 2, PO Box 9600, 2300RC, Leiden, The Netherlands. E.L.Kaijzel@lumc.nl.
authors.forename: Eric L
authors.initials: EL
authors.lastname: Kaijzel
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D015398:Signal Transduction / Q000187:drug effects*; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D023041:Xenograft Model Antitumor Assays
chemical_list: D016328:NF-kappa B; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3; D053179:Caspase 7
keywords: Apoptosis; Bioluminescence; Breast cancer; Gaussia; Multicolor; NF-κB; TNFα