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

Figure 4.

Figure 4

Subcellular distribution of TDP-43 in MNs

(A) Schematics representing the fractionation workflow into nuclear and cytosolic fractions.

(B–F) Immunoblot of nuclear and cytosolic fractions (B) and quantification of nuclear (C) and cytosolic (D) TDP-43 levels, and cytosolic C-terminal fragment of 35 kDa (CTF-35) levels (E and F). Histone H3 (nuclear marker) and actin (cytosolic marker) were used as both loading and fractionation controls. n = 6 extractions from 4 independent differentiations. Pooled data from MNs harvested 4- and 6-week post-plating.

(G) Representative images of MNs differentiated for 6 weeks subjected to immunocytochemistry for TDP-43 (C-terminal antibody) and NF-H. Scale bar, 50 μm.

(H and I) Quantification of TDP-43 distribution using the nuclear/cytosolic ratio of TDP-43 fluorescence signal intensity (H) and the TDP-43/Hoechst correlation coefficient (I). Individual data points represent per-frame mean values from 5 independent experiments. All data shown as mean ± SEM. ∗p < 0.05. Ordinary one-way ANOVA. See also Figure S6.