Titlebook: Hypoxia; Methods and Protocol L. Eric Huang Book 2018 Springer Science+Business Media, LLC 2018 ChIP.CRISPR.Cas9 Gene Editing.VHL binding.R

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Acquisition of Temporal HIF Transcriptional Activity Using a Secreted Luciferase Assay,ition of dynamic and high-throughput data on HIF transcriptional activity during hypoxia and pharmacological activation of HIF. The sensitivity of the assay allows for the secreted luciferase to be consecutively sampled (as little as 1% of the total supernatant) over an extended time period, thus al

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Fluorescence Lifetime Imaging Microscopy (FLIM) as a Tool to Investigate Hypoxia-Induced Protein-Prr fluorophore in close proximity to an appropriate acceptor fluorophore transfers emission energy to the acceptor, resulting in a shorter lifetime of the donor fluorescence. When the respective FRET donor and acceptor are fused with two proteins of interest, a reduction in donor lifetime, as detecte

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Transcriptional Profiling Using RNA-Seq to Study Hypoxia-Mediated Gene Regulation,he transcription factors known as hypoxia-inducible factors (HIFs). HIF1, a heterodimer of hypoxia-stabilized subunit HIF-1alpha and a constitutively expressed subunit HIF-1beta, serves as a key transcription factor that regulates gene expressions which are involved in cell growth, metabolism, and p

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Evaluating the Metabolic Impact of Hypoxia on Pancreatic Cancer Cells, alterations in NAD(P)H/NAD(P). and intracellular ATP levels, and increased utilization of glutamine as the major precursor for fatty acid synthesis. In this chapter, we describe in detail various physiological assays that have been adopted to study the metabolic shift propagated by exposure to hypo

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