Extended Data Fig. 7: SFPQ dissolves FUS condensates in vitro.
From: Small-molecule dissolution of stress granules by redox modulation benefits ALS models
a, Representative images of SFPQ-GFP (top, 10 µM) and FUS-GFP (bottom, 7 µM) protein condensates at a low salt condition (75 mM KCl) and in the presence of H2O2 from >3 independent experiments. b, SDS-PAGE (non-reduced condition) of 10 µM of the purified and untagged SFPQ proteins in diluted state oxidized with the indicated percentages of H2O2 for 30 min. c, Left, representative images of co-incubation of indicated concentrations of SFPQ-GFP and 6 µM of FUS-SNAP at a physiological salt concentration (150 mM KCl) from >3 independent experiments. SFPQ-GFP do not form condensates at 150 mM KCl while they suppress condensation of FUS proteins in dosage-dependent manner. Right, mean ± s.d. of FUS condensate fraction in the presence of GFP (control) or SFPQ-GFP. n = 16 image fields. d, Representative images of FUS-SNAP condensates (4 µM) co-incubated with 40 µM of GFP or SFPQ-GFP at a physiological salt concentration (150 mM KCl) in the presence of indicated percentages of H2O2 from 3 independent experiments. e, Schema of the time course used in Fig. 5c. Cells were firstly cultured in methionine (Met)-free medium and then in each medium (complete medium or Met-free medium supplemented with Met or AHA) before arsenate treatment. f, Left, mean ± s.d. of relative expression levels of SFPQ normalized to those of α-Tubulin in HeLa cells, detected by immunoblotting from 3 independent experiments. Right, mean ± s.d. of nucleus/cytoplasm signal ratio of SFPQ in HeLa cells, detected by immunostaining from 3 independent experiments (n = 363–402 cells). The cells were treated with the indicated medium as in E. p values from Tukey’s test.
