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. 2024 May 16;15:4163. doi: 10.1038/s41467-024-48488-7

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2024

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Fig. 1 — a Volcano plot showing the differential expression analysis (DEA) of RNA sequencing data from brain samples of ALS individuals (n = 323) and normal controls (n = 68). Blue dots: genes upregulated in ALS; red dots: genes downregulated in ALS; gray dots: genes with no statistically significant differences (Differential expression analysis (DEA)). Differentially expressed genes (DEGs) were counted as genes with a False discovery rate (FDR) adjusted p value < 0.05). b, c, e ASRGL1 immunostaining of brain samples. Motor cortex samples (Broadman area 6) of ALS individuals (n = 4), multiple sclerosis individuals (MS) (n = 4), Alzheimer’s disease (AD) (n = 4), and normal controls (NC) (n = 4) were immunostained for ASRGL1. Visual cortex samples (Broadman area 17) from the same ALS individuals were also stained for ASRGL1 and consecutive slides of those samples were stained for TDP-43. ASRGL1+ cells were quantified by automated microscopy and presence of cytoplasmic TDP-43 was analyzed manually in a blinded manner. b Percentage of cells positive for ASRGL1 in FFPE motor cortex samples (BA6), (one-way ANOVA (Mean ± SEM)). c Percentage of ASRGL1+ cells in BA6 and BA17 of the same ALS individuals (two-sided unpaired T-test; Mean ± SEM). d Correlation of the percentage of ASRGL1+ cells and the percentage of cells with cytoplasmic TDP-43 in ALS samples (red dots: BA6; black dots: BA17) (Pearson’s r test; Middle line represents best fit line and error lines represent 95% confidence bands of the best fit line). e Representative images of ASRGL1 immunostaining in brain. f, g Pre-motor cortex samples (BA 6) (20 ALS and 20 normal controls) were analyzed by western blotting to measure ASRGL1 protein (Atlas antibody 029725). f Representative image of a western blot for ASRGL1 and the loading control vinculin. g Levels of ASRGL1 protein (ratio ASRGL1/Vinculin) in ALS brains analyzed by Mann–Whitney test (Median (IQR). h ASRGL1 RNA levels were analyzed in brain samples (BA6) from ALS patients (n = 19) and normal controls (n = 16) by qPCR (Mann–Whitney test (Median (IQR)). i ASRGL1 RNA levels were analyzed in differentiated motor neurons derived from iPSCs lines from ALS individuals (n = 6) and normal controls (n = 6), by qPCR (Mann–Whitney test (Median (IQR)). All pairwise comparisons in the Figure were performed with two-sided tests. Source data are provided as a Source Data file.