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. 2024 Feb 9;27(3):109166. doi: 10.1016/j.isci.2024.109166

Figure 2.

Figure 2

TDP-43 MN cultures form a normal axonal network and maintain viability

(A and C) Representative images of MNs differentiated for 6 weeks subjected to immunocytochemistry for neuronal markers βIII-tubulin (A) and NF-H (C). Scale bar, 100 μm.

(B and D) Quantification of total area and number of branches of βIII-tubulin+ axons (B) and NF-H+ axons (D). n = 3 independent experiments. Two-way ANOVA.

(E) Viability of MN cultures differentiated with and without neurotrophic factors (NF) supplementation over a span of 6 weeks post-plating. n = 4 independent experiments. Two-way ANOVA.

(F–G) Immunoblot (F) and quantification (G) of cleaved caspase 3 (CC3) levels. βIII-tubulin was used as loading control. Extractions were performed in MNs harvested after 6 weeks post-plating. n = 4 independent experiments. Ordinary one-way ANOVA.

(H) Effect of glutamate treatment (0.1 mM glutamate, 24 h) on viability of MNs differentiated for 4 weeks n = 4 independent experiments. Two-way ANOVA.

(I) Effect of ethacrynic acid treatment (50 μM EA, 17 h) on viability of MNs differentiated for 4 weeks. Individual points represent per-well values from 3 independent experiments. Two-way ANOVA. All data shown as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figures S3 and S4.