ArticleNeoplasia (New York, N.Y.)
Green fluorescent protein is a suitable reporter of tumor hypoxia despite an oxygen requirement for chromophore formation.
Article in Neoplasia (New York, N.Y.). The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed.
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- Spatiotemporal analysis of ratiometric biosensors in live multicellular spheroids using SPoRTS.Cell reports methods · 2025Article
- Assessment of Hypoxia in Breast Cancer: Emerging Functional MR Imaging and Spectroscopy Techniques and Clinical Applications.Journal of magnetic resonance imaging : JMRI · 2025Review
- Investigating the effects of stereotactic body radiation therapy on pancreatic tumor hypoxia and microvasculature in an orthotopic mouse model using intravital fluorescence microscopy.Scientific reports · 2024Article
- Interplay between environmental yielding and dynamic forcing modulates bacterial growth.Biophysical journal · 2024Article
- Therapy-induced modulation of tumor vasculature and oxygenation in a murine glioblastoma model quantified by deep learning-based feature extraction.Scientific reports · 2024Article
- Quantitative intravital imaging for real-time monitoring of pancreatic tumor cell hypoxia and stroma in an orthotopic mouse model.Science advances · 2023Article
- Selecting the appropriate indirect viability assay for 3D paper-based cultures: a data-driven study.The Analyst · 2023Article
- Neddylation is required for perinatal cardiac development through stimulation of metabolic maturation.Cell reports · 2023Article
- Lactate-dependent chaperone-mediated autophagy induces oscillatory HIF-1α activity promoting proliferation of hypoxic cells.Cell systems · 2022Article
- Hypoxia-directed tumor targeting of CRISPR-Cas9 and HSV-TK suicide gene therapy using lipid nanoparticles.Molecular therapy. Methods & clinical development · 2022Article
- USP33 deubiquitinates and stabilizes HIF-2alpha to promote hypoxia response in glioma stem cells.The EMBO journal · 2022Article
- Hypoxia and HIF-1 Trigger Marek's Disease Virus Reactivation in Lymphoma-Derived Latently Infected T Lymphocytes.Journal of virology · 2022Article
- Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control.Scientific reports · 2022Article
- Cell line development for continuous high cell density biomanufacturing: Exploiting hypoxia for improved productivity.Metabolic engineering communications · 2021Article
- The MEMIC is an ex vivo system to model the complexity of the tumor microenvironment.Disease models & mechanisms · 2021Article
- Detection of Low Oxygen Microenvironments in a Murine Model of Invasive Pulmonary Aspergillosis Using Pimonidazole.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Serum lipoprotein-derived fatty acids regulate hypoxia-inducible factor.The Journal of biological chemistry · 2020Article
- Fungal biofilm architecture produces hypoxic microenvironments that drive antifungal resistance.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- The focal adhesion protein kindlin-2 controls mitotic spindle assembly by inhibiting histone deacetylase 6 and maintaining α-tubulin acetylation.The Journal of biological chemistry · 2020Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
The oxygen requirement for chromophore formation potentially limits the use of green fluorescent protein as a reporter under hypoxic conditions. In the light of this, the applicability of a hypoxia-responsive enhanced green fluorescent protein (EGFP)-based system to the measurement of tumor hypoxia was tested in human HT 1080 fibrosarcoma cells stably transfected with a destabilized EGFP vector containing the hypoxia-responsive 5HRE-hCMVmp promoter or, as a positive control, the strong constitutive CMV promoter. After various schedules of hypoxia and reoxygenation, EGFP fluorescence of live cells was assessed by flow cytometry, and protein levels were analyzed by Western blot. Fluorescence of CMV promoter positive control cells dropped to 38+/-5% of aerobic levels after 12 hours at <0.02% oxygen, but was unaffected by higher oxygen concentrations. Following 12 hours at <0.02% oxygen, cells transfected with the hypoxia-responsive vector exhibited maximum fluorescence after 4 hours of subsequent reoxygenation, reaching 68+/-2% of the levels in CMV promoter controls under aerobic conditions. With such reoxygenation, these cells exhibited a constant increase in fluorescence between 2% and <0.02% oxygen. EGFP chromophore formation is only affected by near-anoxic oxygen concentrations. The correlation of fluorescence and oxygen concentration is restored by a 4-hour reoxygenation period due to oxidation of pre-synthesized EGFP and a delayed increase in EGFP protein synthesis.
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