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Fig. 2 | Molecular Neurodegeneration

Fig. 2

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

Fig. 2

A315T and Q331K mutants do not prevent the formation of 53BP1 foci in NSC-34 cells Top panels (a) and (b); Confocal microscopy reveals that NSC34-cells expressing wildtype TDP-43 display less DNA damage compared to controls; untransfected (Un) or those expressing mCherry or EGFP only (mCherry, EGFP), determined by (i) total area, (ii) and total volume of 53BP1 foci after treatment with (a) 13.5 μM topoisomerase II inhibitor etoposide or (b) 100 μM H2O2 for 1 h. In contrast, cells expressing ALS-associated mutants A315T and Q331K are not protected from damage compared to wildtype TDP-43. Scale bar 10 μm. Middle panels (a) and (b); Quantification was performed on 3D reconstructions of z-stack images using Imaris software. 2-way ANOVA with Sidak correction for multiple comparison. Mean ± SEM, *p < 0.05, **p<0.01, ***p < 0.001, three independent replicate experiments were performed (etoposide), two independent replicate experiments were performed (H2O2). At least 19 cells/group were analyzed. Bottom panels (a) and (b); Representative 3D reconstruction of confocal images of cells illustrating 53BP1 foci, Scale bar 2 μm

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