Figure 5 | Scientific Reports

Figure 5

From: Two mutations G335D and Q343R within the amyloidogenic core region of TDP-43 influence its aggregation and inclusion formation

Figure 5

Aggregation and inclusion formation of TDP-35 and its mutants.

(a) Immunofluorescence microscopic imaging showing inclusion formation of FLAG-TDP-35 and its mutants in HEK 293T cells. Cells were stained with mouse anti-FLAG (green) and the nuclei were stained with Hoechst (blue). Scale bar = 10 μm. (b) Quantitation of the cells with inclusion bodies in HEK 293T cells overexpressed with FLAG-TDP-35 and its mutants. Cells were counted and the data were statistically analyzed by one-way ANOVA and shown as Mean ± SD (n = 28–30). ***p < 0.001. (c) Supernatant/pellet fractionation assay of the TDP-35 mutants. FLAG-tagged TDP-35 and its mutants were transfected into HEK 293T cells respectively. The cell lysates were fractionated into supernatant (Sup.) and pellet (Pel.) followed by immunoblotting against mouse FLAG and goat actin. (d) Quantification of the amounts of overexpressed TDP-35 species in supernatant and pellet fractions respectively. Acitn was set as a control for relative level. Data were normalized, statistically analyzed by one-way ANOVA and represented as Mean ± SD (n = 3). *p < 0.05; **p < 0.01; n.s., no significance.

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