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. 2020 Sep 25;10(10):1367. doi: 10.3390/biom10101367

Figure 5.

Figure 5

SG assembly mutants impair the formation of TDP-43 cytoplasmic foci in acute and chronic stress. (A) U2-OS (WT) and G3BP1/2ΔΔ cells were unstressed, stressed with 0.5 mM sodium arsenite for 1 h or stressed with 0.5 mM sodium arsenite for 1 h followed by recovery for 2.5 h. Immunofluorescence for TDP-43 utilized the Proteintech 60019-2-Ig antibody, with G3BP staining serving as an SG marker (and conformation of KO status). (B) Endogenous TDP-43 solubility in WT and G3BP1/2ΔΔ cells was determined via differential solubilization using Triton X-100 (TX), Sarkosyl (SS) and SDS (SI). Western blotting was conducted with the Proteintech 10782-2-AP antibody. (C) Quantification of (B). (D) U2-OS (WT) and G3BP1/2ΔΔ cells immunostained as in (A) were left unstressed, stressed with 0.05 mM sodium arsenite for 16 h or stressed with 0.05 mM sodium arsenite for 16 h, followed by recovery for 6 h. Numbers indicated in TDP-43 column indicate average number of TDP-43 foci/cells (see also Supplementary Table S2). Arrowheads indicate cells with TDP-43 foci. Immunofluorescence (IF) was carried out as in (A). (E) Zoom panels indicated by white boxes in (D).