Journal: Molecular Neurodegeneration

Article Title: Impaired nucleocytoplasmic transport in SOD1-mediated ALS

doi: 10.1186/s13024-026-00930-8

Figure Lengend Snippet: SOD1 pathology is associated with disrupted import transport factors and decreased NPC density. a . Immunofluorescence of endogenous RanGAP1 (magenta) in ChAT-positive cells (green) from lumbar spinal cord sections of non-transgenic ( n = 4) and end-stage SOD1 G93A ( n = 4) mice. DAPI (blue) was used to detect the nucleus; scale bar, 20 µm. b . Quantification of the laminar intensity of RanGAP1 represented in ( a ). c . Quantification of the cytosolic distribution of RanGAP1 in spinal motor neurons represented in ( a ). d . Immunofluorescence of endogenous RanGTP (magenta) in ChAT-positive cells (green) from lumbar spinal cord sections of non-transgenic ( n = 3) and end-stage SOD1 G93A ( n = 4) mice. e . Quantification of the RanGTP gradient in spinal motor neurons from staining in ( d ). f . Immunofluorescence of endogenous FG-Nups (recognized with mAb414 antibody ) (magenta) in ChAT-positive cells (green) from lumbar spinal cord sections of non-transgenic ( n = 3) and end-stage SOD1 G93A ( n = 3) mice. g . Inset of the motor neurons nuclear pore complex staining from ( f ) highlighting the diminished staining intensity and the presence of FG-Nups inclusions in spinal cords of SOD1 G93A mice compared to control; scale bar, 5 µm h . Quantification of the laminar intensity of FG-Nups represented in ( f ). i . Pie chart of the number of motor neurons with puncta in ( f ). Graphs represent quartiles (boxes), 50th percentiles (center lines) and range (10–90; whiskers). Four independent experiments for ( b ) and ( c ) (dots represent quantified motor neurons from all experiments. Rank-based two-samples t-test, p-values adjusted for clustering; * p < 0.05, *** p < 0.001), three independent experiments for e (dots represent quantified motor neurons from all experiments. Rank-based two-samples t-test, p-values adjusted for clustering; * p < 0.05), three independent experiments for ( h ) (dots represent quantified motor neurons from all experiments. Rank-based two-samples t-test, p-values adjusted for clustering; *** p < 0.001)

Article Snippet: SC-28322 , Santa-Cruz , 1:100 , RanGAP1 (C-5), mouse.

Techniques: Immunofluorescence, Transgenic Assay, Staining, Control

Journal: Molecular Neurodegeneration

Article Title: Impaired nucleocytoplasmic transport in SOD1-mediated ALS

doi: 10.1186/s13024-026-00930-8

Figure Lengend Snippet: Nucleocytoplasmic transport impairment in fibroblasts carrying SOD1 mutation. a . Representative confocal images of immortalized fibroblasts from SOD1-ALS patients and healthy controls immunostained for FG-Nups (gray) and counterstained with DAPI (blue). Scale bar, 20 µm. b . Quantification of nuclear FG-Nups intensity in six mutant SOD1-ALS fibroblasts (five SOD1–A4V, one SOD1–D90A) and six controls. The box plot reflects median per well; each data point indicates the average for each line from two independent experiments. Statistical significance was assessed across data points. Unpaired t-test; ** p < 0.01. c . Confocal imaging after immunofluorescence staining of RanGAP1 (green) and DAPI staining (blue). Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu $$\end{document} m. d . Quantification of RanGAP1 accumulation at the nuclear membrane (laminar intensity) from six healthy controls and six SOD1-ALS patients (five SOD1–A4V and one SOD1–D90A) in ( c ). The box plot reflects the median per well; each data point indicates the average per line. Statistical significance was assessed across data points. Unpaired t-test; * p < 0.05. e . Inset of the fibroblasts RanGAP1 staining from ( c ) highlighting the presence of RanGAP1 inclusions in fibroblasts carrying SOD1 mutants (A4V, D90A) compared to control; scale bar, 3 µm. f . Quantification of RanGAP1 cytoplasmic puncta in fibroblasts using confocal imaging. The box plot reflects the median per well, and each data point indicates the average for each line from three individual experiments. Statistical significance was assessed across data points. Unpaired t-test; * p < 0.05. g. Flow cytometry-based imaging of fibroblasts stained for RanGAP1 (green) and DAPI (blue). Bright field is shown in gray. Scale bar, 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu $$\end{document} m. h . Flow cytometry-based imaging analysis of cytosolic RanGAP1 intensity in fibroblasts from two healthy controls, one SOD1–A4V, and one SOD1–D90A ALS patients. Each dot represents the average from a single experiment (three experiments per line); squares indicate data from the SOD1–D90A line. Statistical analysis was performed using an unpaired t-test comparing the experimental averages for each line; ** p < 0.01

Article Snippet: SC-28322 , Santa-Cruz , 1:100 , RanGAP1 (C-5), mouse.

Techniques: Mutagenesis, Imaging, Immunofluorescence, Staining, Membrane, Control, Flow Cytometry

Journal: Molecular Neurodegeneration

Article Title: Impaired nucleocytoplasmic transport in SOD1-mediated ALS

doi: 10.1186/s13024-026-00930-8

Figure Lengend Snippet: Nucleocytoplasmic transport disruption in SOD1-ALS postmortem spinal cord tissues. a . Confocal imaging after immunofluorescence staining of RanGAP1 (green), B8H10 for misfolded SOD1 (magenta), ChAT (cyan), and DAPI (blue) in postmortem lumbar spinal cord from a control individual and a patient with SOD1 A4V mutation. Scale bar, 50 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu $$\end{document} m. b . Quantification of RanGAP1 accumulation at the nuclear membrane (laminar intensity) in lumbar spinal cords from controls ( n = 5) and SOD1 A4V ALS patients ( n = 4). c . Confocal imaging after immunofluorescence staining of FG-Nups (magenta) and DAPI (blue) in postmortem cervical spinal cord from a control individual and a patient carrying SOD1 A4V mutation. Scale bar, 20 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu $$\end{document} m. d . Quantification of FG-Nups intensity at the nuclear membrane as the ratio of nuclear envelope to nucleoplasm signal intensity in cervical spinal cord sections from controls ( n = 3) and SOD1 A4V ALS patients ( n = 3). e . Confocal imaging after immunofluorescence staining of XPO1 (magenta) and DAPI (blue) in postmortem cervical spinal cord from a control individual and a patient carrying SOD1 A4V mutation. Scale bar, 20 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu $$\end{document} m. f . Quantification of cytosol-to-nucleus ratio of XPO1 signal intensity in cervical spinal cord sections from controls ( n = 3) and SOD1 A4V ALS patients ( n = 3). Graphs represent quartiles (boxes), 50th percentiles (center lines) and range (10–90; whiskers. For ( b ) data were quantified from 5 healthy controls and 4 SOD1 A4V ALS patients (dots represent quantified motor neurons from all experiments. Rank-based two-samples t-test, p-values adjusted for clustering; *** p < 0.001). For ( d ) and ( f ), data were quantified from 3 healthy controls and 3 SOD1 A4V ALS patients (dots represent quantified motor neurons from all experiments. Unpaired parametric t-test, ** p < 0.01, *** p < 0.001)

Article Snippet: SC-28322 , Santa-Cruz , 1:100 , RanGAP1 (C-5), mouse.

Techniques: Disruption, Imaging, Immunofluorescence, Staining, Control, Mutagenesis, Membrane

Journal: bioRxiv

Article Title: SUMO E1 covalent allosteric inhibitors modulate polyamine synthesis via the MAT2A-AdoMetDC axis

doi: 10.1101/2024.12.12.627095

Figure Lengend Snippet: The new SUMO E1 CAIs perturb the SUMOylation pathway. T47D cells were treated with MCULE-3064932370 and MCULE-6830015021 for 48 h before evaluation. (a) The cellular levels of RanGAP1, RanGAP1-SUMO, and SUMO1 were assessed by Western blotting. (b) The cellular levels of Uba2 and Ubc9 were assessed by Western blotting.

Article Snippet: The rabbit monoclonal anti-RanGAP1 (Cat. NO. R383099) and anti-MAT2A (Cat. NO. R389369) antibodies were from ZenBio, Inc. (U.S.A.).

Techniques: Western Blot