Fig. 1: Localization of endogenous wild type (WT) and mutant FUS to mitochondria: Both ALS associated FUS pathology or glucose oxidase stress increased mitochondrial localization.

a, b Immunoblots (IB) of cellular fractionization showing FUS in mitochondria. Total, cytoplasm and mitochondrial extracts are isolated from HEK 293 WT cells with and without glucose oxidase (GO) 100 ng/ml for 1 h and from FUS knockout (KO) cells in a. b corresponds to patient-derived iPSC differentiated into motor neurons with and without GO treatment, the data are presented as mean ± s.e.m. from three individual experiments, with the individual data points shown. c–e show proximity ligation assay (PLA) performed in patient derived fibroblasts and motor neurons, c corresponds to PLA of FUS with Tom20 without any stress along with quantifications. Scale bar = 20 µM. Box plot parameters: WT (minima 9, maxima 51, median 24, 25^th percentile is 14 and 75^th percentile is 39). For R521H (minima 51, maxima 125, median 89, 25^th percentile is 115 and 75^th percentile is 216). For P525L (minima 148, maxima 272, median 218, 25^th percentile is 189 and 75^th percentile is 239). d represents the comparison between PLA foci for FUS and HSP60 with and without GO and their quantifications. Box plot parameters: FUS WT (minima 8, maxima 21, median 12, 25^th percentile is 10 and 75^th percentile is 15). For FUS WT GO (minima 23, maxima 54, median 36, 25^th percentile is 31 and 75^th percentile is 42). For FUS R521H (minima is 13, maxima 37, median is 27, 25th percentile is 21 and 75th percentile is 32). For R521H GO, (minima 41, maxima 79, median 56, 25th percentile is 47 and 75th percentile is 69). For FUS P525L, (minima 34, maxima 72, median 51, 25th percentile is 41 and 75th percentile is 65). For FUS P525L GO (minima 52, maxima 85, median 64, 25th percentile is 57 and 75th percentile is 74). e corresponds to PLA between FUS and TOM20 in motor neurons with and without GO treatment Nuclei stained with DAPI and mito-tracker green is used to counter stain mitochondria. Scale bar = 20 μM. The data are presented as mean ± s.e.m. from three individual experiments, quantification of PLA foci was derived from 25 cells. FUS WT (minima 0, maxima 4, median 1, 25^th percentile is 1 and 75^th percentile is 2), for FUS WT GO (minima 2, maxima 8, median 4, 25^th percentile is 3 and 75^th percentile is 5). For FUS R521H (minima 1, maxima 9, median 4, 25th percentile is 2 and 75th percentile is 5). For R521H GO (minima 5, maxima 16, median 9, 25th percentile is 7 and 75th percentile is 12). For FUS P525L (minima 1, maxima 8, median 4, 25th percentile is 3 and 75th percentile is 6). For FUS P525L GO (minima 4, maxima 19, median 13, 25th percentile is 9 and 75th percentile is 16). All statistical analysis were performed by two-sided student-t test using graph pad prism software. Source data are provided as a Source Data file.