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. 2023 Jul 31;32(20):2966–2980. doi: 10.1093/hmg/ddad122

Figure 3.

Figure 3

TDP-43 aggregates are more soluble with ORF57. Experiments conducted in mCherry and ORF57 mCherry expressing SH-SY5Y TDP-43ΔNLS cells with SA stress. (A) Western blot of TDP-43ΔNLS::GFP RIPA-soluble and in-soluble fractions. (B) Quantification of %RIPA-Soluble TDP-43ΔNLS::GFP calculated as soluble TDP-43 divided by the sum of soluble and insoluble TDP-43. (C) Quantification of %RIPA-insoluble TDP-43ΔNLS::GFP. (D) Time course experiment measuring TDP-43ΔNLS::GFP aggregate recovery at 15, 30 min, 1 , 2 , 3.5 and 6.5 h after SA removal. (E) Imaging of TDP-43ΔNLS using FRAP. (F) Quantification of D with exponential one-phase decay best fit curves. (G) Analysis of TDP-43ΔNLS mobility using FRAP. mCherry controls cells had a mobile fraction of 0.03 and a t1/2 of 111 s; ORF57::mCherry expressing cells had a higher mobile fraction of 0.2 and a t1/2 of 68 s. Unpaired t-tests, *P < 0.05. Scale bars 20 μm.