FIGURE

Fig. 4

ID
ZDB-FIG-220921-80
Publication
Chaytow et al., 2022 - Targeting phosphoglycerate kinase 1 with terazosin improves motor neuron phenotypes in multiple models of amyotrophic lateral sclerosis
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Fig. 4

Terazosin protects against NaArO2-induced cell death, increases glycolysis and decreases mitochondrial respiration in mESC-MNs. (a) TDP-43M337V mESC-MNs show a decreased survival in response to sodium arsenite (NaArO2) stress compared to unstressed controls. Survival following terazosin treatment at all concentrations is maintained at 100%. (n = 3 differentiations; One-way ANOVA P < 0.001 with Dunnett's multiple comparisons). (b) Traces from the Seahorse analyser glycolytic rate assay showing glycolytic proton efflux rate (GlycoPER) following mitochondrial inhibition (oligomycin) and inhibition of the glycolysis pathway (2-deoxy-D-glucose; 2-DG). (c) TDP-43M337V mESC-MNs demonstrate a significantly lower rate of basal glycolysis compared to TDP-43WT controls, which is rescued by treatment of 2.5 µM terazosin (Tz) (One-way ANOVA P = 0.017 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P = 0.034, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P > 0.99). (d) TDP-43M337V mESC-MNs show significantly reduced rates of compensatory glycolysis compared to TDP-43WT controls, which is rescued by treatment of 2.5 µM terazosin (Tz) (One-way ANOVA P = 0.055 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P = 0.038, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P = 0.22). (e) Traces from the Seahorse analyser showing oxygen consumption rate (OCR) following mitochondrial inhibition (oligomycin), mitochondrial uncoupling (FCCP) and electron transport chain inhibition (antimycin A/rotenone). (f) Treatment with 2.5 µM terazosin significantly decreases basal respiration in TDP-43M337V mESC-MNs compared to TDP-43WT controls (One-way ANOVA P = 0.001 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P > 0.99, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P = 0.003). (g) Treatment with 2.5 µM terazosin significantly decreases maximal respiration in TDP-43M337V mESC-MNs compared to TDP-43WT controls (One-way ANOVA P = 0.003 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P = 0.091, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P = 0.002). (h) Treatment with 2.5 µM terazosin significantly decreases ATP production in TDP-43M337V mESC-MNs compared to TDP-43WT controls (One-way ANOVA P = 0.004 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P = 0.87, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P = 0.018). (i) Treatment with 2.5 µM terazosin does not change the spare respiratory capacity in TDP-43M337V mESC-MNs compared to TDP-43WT controls (One-way ANOVA P = 0.19 with Bonferroni's multiple comparison test: TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2P = 0.16, TDP-43WT + NaArO2 vs TDP-43M337V + NaArO2 + 2.5 µM Tz P = 0.38). Each data point represents a separate differentiation; n = 7 per glycolysis analysis; n = 6 per respiration analysis. Error bars represent s.e.m., ns=non-significant, * = P < 0.05, ** = P < 0.01.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
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