Extended Data Fig. 1: HeLa cell and in vitro follow-up screen identifying lipoamide.
From: Small-molecule dissolution of stress granules by redox modulation benefits ALS models
a, Workflow for screening small molecules for effects on FUS–GFP localization in HeLa cells ex vivo. b, Ranked Mahalanobis distances for all 1600 compounds screened (mean from six fields of view) where high values indicate more compound effect. Several automated measures of FUS localization were combined into a single Mahalanobis distance score; the largest contributors were cytoplasmic FUS condensate number and area (see the method section). A cut-off of 130 was used to select 47 compounds for further analysis. c, The subcellular localization of FUS–GFP in unstressed HeLa cells, stressed cells with compound solvent (DMSO) negative control, and with the positive control emetine, as observed in the screen in B. Stress causes nuclear export of FUS and formation of stress granules (cytoplasmic liquid FUS-containing condensates). Insets, magnified images in the boxed areas. d, Workflow for screening small molecules for effects on FUS condensation of purified FUS–GFP in vitro. e, Ranked Z scores of change in condensate droplet number and signal partition into FUS–GFP droplets (formed under low salt conditions) where larger positive or negative values mean more compound effect. Scores were calculated at the maximum concentration at which the compound solvent (DMSO) negative control had no significant effect; 100 μM. Lipoamide, surfactant and heterotri-/tetracyclic compounds are indicated by data point colour. f, Appearance of the droplets with compound solvent (DMSO) negative control or examples of compound classes: cetylpyridinium chloride (surfactant), lipoamide or mitoxantrone (heterotricyclic). Note the larger drops with cetylpyridinium chloride and lipoamide and the fewer smaller drops with mitoxantrone.
