Immunodetection of Human Double Homeobox 4

Overview

Journal Hybridoma (Larchmt)

Date 2011 May 3

PMID 21529284

Citations 38

Authors

Linda N Geng

Ashlee E Tyler

Stephen J Tapscott

Affiliations

Soon will be listed here.

Abstract

Double homeobox 4 (DUX4) is a candidate disease gene for facioscapulohumeral dystrophy (FSHD), one of the most common muscular dystrophies characterized by progressive skeletal muscle degeneration. Despite great strides in understanding precise genetics of FSHD, the molecular pathophysiology of the disease remains unclear. One of the major limitations has been the availability of appropriate molecular tools to study DUX4 protein. In the present study, we report the development of five new monoclonal antibodies targeted against the N- and C-termini of human DUX4, and characterize their reactivity using Western blot and immunofluorescence staining. Additionally, we show that expression of the canonical full coding DUX4 induces cell death in human primary muscle cells, whereas the expression of a shorter splice form of DUX4 results in no such toxicity. Immunostaining with these new antibodies reveals a differential effect of two DUX4 isoforms on human muscle cells. These antibodies will provide an excellent tool for investigating the role of DUX4 in FSHD pathogenesis.

Citing Articles

Facioscapulohumeral Dystrophy: Molecular Basis and Therapeutic Opportunities.

Arends T, Hamm D, van der Maarel S, Tapscott S Cold Spring Harb Perspect Biol. 2024; .

PMID: 39009417 PMC: 11733064. DOI: 10.1101/cshperspect.a041492.

Human DUX4 and mouse Dux interact with STAT1 and broadly inhibit interferon-stimulated gene induction.

Spens A, Sutliff N, Bennett S, Campbell A, Tapscott S Elife. 2023; 12.

PMID: 37092726 PMC: 10195082. DOI: 10.7554/eLife.82057.

The double homeodomain protein DUX4c is associated with regenerating muscle fibers and RNA-binding proteins.

Claus C, Slavin M, Ansseau E, Lancelot C, Bah K, Lassche S Skelet Muscle. 2023; 13(1):5.

PMID: 36882853 PMC: 9990282. DOI: 10.1186/s13395-022-00310-y.

Proximity ligation assay to detect DUX4 protein in FSHD1 muscle: a pilot study.

Beermann M, Homma S, Miller J BMC Res Notes. 2022; 15(1):163.

PMID: 35538497 PMC: 9092897. DOI: 10.1186/s13104-022-06054-8.

Downstream events initiated by expression of FSHD-associated DUX4: Studies of nucleocytoplasmic transport, γH2AX accumulation, and Bax/Bak-dependence.

Masteika I, Sathya A, Homma S, Miller B, Boyce F, Miller J Biol Open. 2022; 11(2).

PMID: 35191484 PMC: 8890089. DOI: 10.1242/bio.059145.

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