Extended Data Fig. 2: Lipoamide characterization in HeLa cells.
From: Small-molecule dissolution of stress granules by redox modulation benefits ALS models
a, Mean ± s.d. of percentage of HeLa cells with ≥ 3 G3BP1-positive stress granules (SGs). Cells were treated with 1 mM arsenate for 1 h to induce SGs, followed by 10 µM lipoamide, lipoic acid, or 0.1% DMSO (control) in the presence of arsenate for indicated minutes. n = 52–248 cells from 3 independent experiments. b, Mean ± s.d. of relative intensity levels of puromycin normalized to those of GAPDH in HeLa cells treated with 10 µM lipoamide (0.1% DMSO as the control) for 1 h, followed by 1 mM arsenate for 1 h and 91.8 µM puromycin for 5 min, detected by immunoblotting from 3 independent experiments. c, Images of HeLa cells expressing GFP-tagged markers of other membrane-less organelle compartments subjected to 1 h treatment with 10 μM lipoamide (or DMSO control). Where unclear, thep position of nuclei is indicated with a broken outline. Lipoamide does not disrupt P bodies (DCP1A), Cajal bodies (COIL), or DNA damage foci (TRP53BP1). d, Images of U2OS cells stained with indicated markers of other membrane-less subcellular compartments subjected similarly to HeLa cells in G. Lipoamide also did not disrupt nuclear speckles (SC35), PML bodies (SP100), nucleoli (NPM1), or heterochromatin (HP1α). e, Images of HeLa cells expressing FUS–GFP subjected to different stresses – arsenate, sorbitol (osmotic), heat, or 6-deoxyglucose (DG; glycolysis) – with concurrent treatment with 10 μM lipoamide or lipoic acid. f, HeLa cells were treated with 0.1% DMSO (control), 10 µM lipoamide, or indicated concentrations of other known and potential antioxidants (ascorbic acid as representative images on the top) for 1 h, followed by 1 mM arsenate for 1 h in the presence of each chemical. SGs were labeled with G3BP1. Bottom, percentage of HeLa cells with ≥ 3 G3BP1-positive SGs from n = 119–202 cells.
