Fig. 1. TSP-1 inhibits Aβ-induced mitochondrial fission in HT22 cells.

a HT22 cells were stained with MitoTracker green dye to visualize mitochondrial morphology. Magnified images in lower panels are the areas marked with white lines. Scale bar, 20 μm. b, c Mitochondrial length, represented as aspect ratio and mitochondrial circular shape, represented as form factor were analyzed with the Image J program software. TSP-1 treatment significantly recovered Aβ-induced mitochondrial fragmentation. Data were obtained from at least three replicates for each group, and 100–150 cells were analyzed for each group. Data are presented as mean ± SEM. *p < 0.05, ***p < 0.001 vs. vehicle (DMSO)-treated cells; ^#p < 0.05 vs. Aβ-treated cells. d The effect of TSP-1 on mitochondrial morphology was analyzed in EM images. Arrows indicate normal mitochondria. Arrow heads indicate fissioned mitochondria. Dashed circle indicates damaged (void) mitochondria. Scale bar, 1 μm. e–g Average mitochondrial length, number of damaged (void) mitochondria per one cell, and the total number of mitochondria per one cell were quantified. TSP-1 treatment significantly recovered mitochondrial length, decreased number of damaged mitochondria, and the total number of mitochondria per one cell. Data were obtained from at least two replicates, and more than total 10 mitochondria were analyzed for each group. Data are presented as mean ± SEM. **p < 0.01, ***p < 0.001 vs. vehicle (DMSO)-treated cells; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 vs. Aβ-treated cells