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Fig. 5 | Molecular Neurodegeneration

Fig. 5

From: Impaired NHEJ repair in amyotrophic lateral sclerosis is associated with TDP-43 mutations

Fig. 5

TDP-43 is recruited to 53BP1 DNA repair foci. Super-resolution imaging of NSC-34 cells expressing mCherry-tagged wildtype, Q331K, A315T TDP-43, or empty vector, enabled quantification of sub-diffraction colocalization between TDP-43 and 53BP1, in cells treated with 13.5 μM etoposide for 1 h. a Quantification of TDP-43/53BP1 co-localisation using the Interaction Factor ImageJ plugin, which was developed specifically for nuclear SR imaging. b-e. Representative SR images of etoposide-treated cells expressing mCherry-tagged wildtype, mutant Q331K, or A315T TDP-43, or empty vector, and immunostained for 53BP1. Low resolution epifluorescence DAPI images were acquired for nuclear localisation. Zoom-in regions of the boxed areas show prominent 53BP1 foci and mCherry-TDP-43 colocalization. t-test between wildtype compared to A315T, Q331K, and mCherry only, ***p < 0.001. Scale bars 3 μm in whole cell images, 250 nm in zoomed regions, N ≥ 30. (f) Immunoprecipitation of GFP Trap of NSC-34 cells expressing wildtype, Q331K, A331T TDP-43 or EGFP only. Wildtype and TDP-43 mutants co-immunoprecipitate with 53BP1, but not with control EGFP

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