Journal: Acta Neuropathologica
Article Title: TDP-43 impairs glycolysis by sequestering hexokinase 1 in amyotrophic lateral sclerosis
doi: 10.1007/s00401-026-02996-6
Figure Lengend Snippet: Pathological TDP-43 elicits glycolytic impairment in cells. a , b Agilent seahorse glycolytic stress test was performed on n = 3 independent biological repeats. Glycolysis and Glycolytic Capacity were measured according to manufacturer’s protocol. ECAR values were normalized to total protein then plotted against time. Individual glycolysis and glycolytic capacity values were normalized to total protein level then to individual control from each biological repeat. a Stable HEK293 cells were overexpressed with either YFP (control), TDP-43 WT or TDP-43 ΔNLS . Data were analyzed by one-way ANOVA followed by post hoc Tukey’s test. b TDP-43 G298S patient iPSC and control iPSC were differentiated into motor neurons with our established protocol. On day 26 following differentiation, cells were treated with MG-132 (1 μM) for 24 h. Data were analyzed by Student’s t-test. c YFP (control), TDP-43 WT or TDP-43 ΔNLS stable cells were transiently transfected with various glycolysis shRNAs. Cell viability was measured via MTT assay. Decreased cell viability in all TDP-43 WT and TDP-43 ΔNLS with various glycolysis shRNA (n = 3). Data were analyzed by one-way ANOVA followed by post hoc Tukey’s test. All data are mean ± SE
Article Snippet: The following constructs pcDNA3.2-YFP (addgene #84910), pcDNA3.2-TDP-43 WT -YFP (human) (addgene #84911), pcDNA3.2-TDP-43 ΔNLS -YFP (human) (addgene #84912), pLD-puro-Cc-TARDBP-A315T_VA Plasmid (human TDP-43 A315T) (addgene #141329), pLD-puro-Cc-TARDBP-WT_VA Plasmid (human TDP-43 WT) (addgene #141327), and shRNA control (addgene: #8453) were purchased from addgene.
Techniques: Control, Transfection, MTT Assay, shRNA
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