Fig. 1: Attachment of SUMO2 protects TDP-43 from stress-induced aggregation.
From: Induced proximity to PML protects TDP-43 from aggregation via SUMO–ubiquitin networks
a, Schematic representation of the solubility assay. HEK293T cells transiently expressing Flag-TDP-43, Flag-SUMO2–TDP-43 or Flag-4×SUMO2–TDP-43 were lysed in an NP-40-containing lysis buffer. Before lysis, cells were exposed to HS (43 °C, 1 h), SA (0.5 mM, 1 h) or left untreated. Lysates were fractionated into an NP-40-insoluble pellet and an NP-40-soluble supernatant fraction and analyzed by western blotting. Panel a is created with BioRender.com. b, Solubility assay. HEK293T cells expressing Flag-TDP-43, Flag-SUMO2–TDP-43 or Flag-4×SUMO2–TDP-43 for 48 h were treated with SA (0.5 mM, 1 h) or left untreated before lysis and fractionation (as described in a). NP-40-soluble/NP-40-insoluble fractions are indicated by green and red bars. Statistical analysis of three independent replicates is provided in Extended Data Fig. 1a. c, As in b, but cells were subjected to HS (43 °C, 1 h) or left untreated. Statistical analysis of three independent replicates is provided in Extended Data Fig. 1a. d, As in b and c, but cells were additionally treated with MG132 (10 µm, 4 h) or DMSO. Statistical analysis of three independent replicates is provided in Extended Data Fig. 1c. e, As in b–d, but cells were treated with DBeQ (20 µm, 4 h) or DMSO. Statistical analysis of three independent replicates is provided in Extended Data Fig. 1d. HS, heat stress; SA, sodium arsenite.
