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. 2019 Dec 27;9:19928. doi: 10.1038/s41598-019-56483-y

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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TDP-43 forms inclusions within micronuclei induced by metabolic stress which co-aggregate with endogenous RGNEF in HEK293T cells. (A,B) 3D reconstruction from confocal set of images of a HEK293T cell showing TDP-43 aggregates within a micronucleus after cellular metabolic stress using lactate. (A) Shows xy axis view and (B) shows z axis view of the cell. MN = micronucleus and N = nucleus. (C) Co-aggregation between endogenous TDP-43 and endogenous RGNEF within a micronucleus after metabolic stress in HEK293T cells (white arrows). (D,E) Absence of aggregate formation for FUS/TLS (D) and sirtuin 1(SIRT1) (E) within micronuclei in HEK293T cells. TDP-43 inclusions are indicated (white arrows). (F) TDP-43 aggregates within micronuclei in HEK293T after metabolic stress cells are immunoreactive for phospho(409/410)-TDP-43 antibody which only stain insoluble inclusions. (G) 3D reconstruction from confocal set of images of the cell in (F) showing the phospo(409/410)-TDP-43 positives aggregates. (H) Control HEK293T cells showing absence of immunoreactive for phospho(409/410)-TDP-43 antibody. Immunofluorescence primary antibodies used were anti-TDP-43, anti-RGNEF, anti-FUS/TLS, anti-sirtuin 1 (SIRT1) and anti-NPCP. Hoechst was used as nucleic acid staining. Scales are indicated in the images.