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Issue 39, 2014
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Single-molecule quantification of lipotoxic expression of activating transcription factor 3

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Abstract

Activating transcription factor 3 (ATF3) is a member of the mammalian activation transcription factor/cAMP, physiologically important in the regulation of pro- and anti-inflammatory target genes. We compared the induction of ATF3 protein as measured by Western blot analysis with single-molecule localization microscopy dSTORM to quantify the dynamics of accumulation of intranuclear ATF3 of triglyceride-rich (TGRL) lipolysis product-treated HAEC (Human Aortic Endothelial Cells). The ATF3 expression rate within the first three hours after treatment with TGRL lipolysis products is about 3500 h−1. After three hours we detected 33 090 ± 3491 single-molecule localizations of ATF3. This was accompanied by significant structural changes in the F-actin network of the cells at ∼3-fold increased localization precision compared to widefield microscopy after treatment. Additionally, we discovered a cluster size of approximately 384 nanometers of ATF3 molecules. We show for the first time the time course of ATF3 accumulation in the nucleus undergoing lipotoxic injury. Furthermore, we demonstrate ATF3 accumulation associated with increased concentrations of TGRL lipolysis products occurs in large aggregates.

Graphical abstract: Single-molecule quantification of lipotoxic expression of activating transcription factor 3

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Publication details

The article was received on 22 Jul 2014, accepted on 28 Aug 2014 and first published on 29 Aug 2014


Article type: Paper
DOI: 10.1039/C4CP03260H
Citation: Phys. Chem. Chem. Phys., 2014,16, 21595-21601
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    Single-molecule quantification of lipotoxic expression of activating transcription factor 3

    I. Yahiatène, H. H. Aung, D. W. Wilson and J. C. Rutledge, Phys. Chem. Chem. Phys., 2014, 16, 21595
    DOI: 10.1039/C4CP03260H

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