Figure 5: The effects of RNAs or TDP-43 N terminus on intrinsic propensity of TDP-43 C terminus. | Nature Communications

Figure 5: The effects of RNAs or TDP-43 N terminus on intrinsic propensity of TDP-43 C terminus.

From: The self-interaction of native TDP-43 C terminus inhibits its degradation and contributes to early proteinopathies

Figure 5

(a) GFP-tagged mTDP-43 FL and the mutant F147, 149L (RD) were transfected into 293T cells. Compared with cells transfected with FL, mutant transfectants showed a 20-fold increase in granule formation. The results of three independent experiments were analysed. All data are presented as means with s.d. (n=3). *P<0.05 by Student's t-test. (b) An examination of TDP-43 proteins in presence of RNase. The protein lysate of 293T cells was used for immunoprecipitation with anti-TDP-43 antibody and was further examined by immunoblotting with anti-TDP antibody. Dimers of TDP were increased in presence of RNase. (c) Western blotting analysis of the solubility of GFP-tagged mutant F147, 149L. (d) The truncated mTDP-43 variants containing PLD formed aggregates in 293T cells. A scheme of the TDP-43 construct, top, illustrates functional domains. NΔ, IIPLD and NPLD, but not PLDΔ, formed aggregates in the nucleus or cytosol. Amorphous mTDP-43-NPLD (the C terminus fused with the N terminus) proteins frequently formed doughnut-shaped aggregates. Scale bars represent 10 μm. (e) Validation of the solubility of GFP-tagged mTDP-43 variants by western blot; *, expected molecular weight. (f) Examination of the toxicity of TDP-43 aggregates. The 293T cells were transfected with GFP-tagged variants of mTDP-43. GFP-positive cells were counted at 24, 48 and 72 h post-transfection. All data are presented as means with s.d. (n=5). *P<0.05 by analysis of variance.

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