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Acquisition of Temporal HIF Transcriptional Activity Using a Secreted Luciferase Assay
Miguel A S Cavadas1,2,3, Cormac T Taylor1,2, Alex Cheong4,5,6
1Systems Biology Ireland, University College Dublin, Dublin, Ireland.
Abstract:
Here we describe a simple method based on secreted luciferase driven by a hypoxia-inducible factor (HIF) response element (HRE) that allows the acquisition 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 allowing the acquisition of time-resolved HIF transcriptional activity.
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