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. 2022 Feb 2;11(3):516. doi: 10.3390/cells11030516

Figure 1.

Figure 1

TDP-43 species were differently distributed in cells, LVs and SVs. (A,B) Representative nanoparticle-tracking analysis (NTA) distribution of LVs (A) and SVs (B) extracted from NSC-34 cells. (A,B) x-axis = vesicles dimension, expressed in nm; y-axis = vesicles concentration, expressed as number of particles/mL. (C,D) Representative transmission electron microscopy (TEM) of NSC-34 cells-derived LVs (C) and SVs (D). Scale bar: 200 nm. (E) The graph represents the mean (left graph) and the mode (right graph) size ± SD of LVs (200.9 ± 40.7 nm; 136.1 ± 14.1 nm, respectively) and SVs (117.2 ± 40.3 nm; 96.0 ± 40.5 nm, respectively) of n = 3 biological replicates analyzed by NTA (* p < 0.05, unpaired one-tailed t-test). (F) Representative WB analysis of EVs markers in NSC-34-derived LVs and SVs samples. (G–J) WB analysis of TDP-43, TDP-35 and TDP-25 in total (i.e., soluble + insoluble) RIPA-extracted proteins from NSC-34 cells (C), LVs and SVs released in basal conditions (G) and relative quantifications (H–J). The bar graphs represent the relative abundance of TDP-43, TDP-35 and TDP-25 in cells (H), LVs (I) and SVs (J), expressed as mean [(optical density of a single TDP-43 species/optical densities of all the immunoreactive TDP-43 species) × 100] ± SD of n = 3 biological replicates. (K–N) WB analyses of RIPA-soluble (left-panels) and RIPA-insoluble (right-panels) fractions of NSC-34 cells, LVs and SVs (K) and relative quantifications (L–N). The bar graphs represent the relative abundance of each TDP-43 species in the soluble or insoluble fraction of cells (L), LVs (M) and SVs (N), expressed as [(optical density of soluble or insoluble/optical densities of soluble + insoluble TDP) × 100] ± SD of n = 3 biological replicates. (L) Cells: TDP-43, 72.8 ± 5.4% soluble; TDP-35, 94.3 ± 2.4% soluble; TDP-25, 96.0 ± 2.7% soluble. (M) LVs: TDP-43, 85.6 ± 13.8% insoluble; TDP-35, 96.9 ± 3.0% insoluble; TDP-25, 96.9 ± 3.8% insoluble. (N) SVs: TDP-43, 76.0 ± 19.1% insoluble; TDP-35, 83.7 ± 3.8% insoluble; TDP-25, 65.4 ± 7.8% insoluble.