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. 2018 May 4;8:7083. doi: 10.1038/s41598-018-25008-4

Figure 8.

Figure 8

Addition of a bulky tag on TDP-43 disrupts nuclear export. (a) Example heterokaryons from a variation of the heterokaryon shuttling assay. HeLa cells were co-transfected with Flag TDP-43 and tdTomato TDP-43 and shuttling assay was performed as in Fig. 3. Heterokaryons were identified by presence of Flag TDP-43 within a recipient nucleus. Flag TDP-43 was visualized via immunofluorescence with Flag antibody, tdTomato TDP-43 was visualized via direct fluorescence. Nuclei were detected using Hoechst stain. Images are representative of 4 independent experiments. Recipient nuclei are indicated by arrow. Scale bar- 10um. (b) Quantification of shuttling assay as in 1b. Results are pooled from 4 independent experiments. hnRNPC- 15 heterokaryons counted. Cotransfected with Flag TDP-43 and tdTomato TDP-43–28 heterokaryons counted. * indicates significant difference between groups, p < 0.001. To compare hnRNPC vs Flag TDP-43, and hnRNPC vs tdTomato TDP-43, Mann Whitney rank sum test was used. To compare Flag TDP-43 vs tdTomato TDP-43, Wilcoxon signed rank test was used.