Figure 7.

ART derivatives, but not control drug GCV, induce significant dose-dependent shortening of mitochondria in primary human fibroblasts. HFFs were treated with the indicated concentrations of compounds or remained untreated for 3 d, after which they were fixed and stained using DAPI and antibody against mitochondrial marker protein prohibitin. (A) Treatment with control drug GCV did not change mitochondrial architecture, as shown by no significant alteration to mean mitochondrial length. (B) Treatment with 10 µM of ART resulted in a statistically significant shortening of mitochondria. (C) Treatment with 10 µM as well as 1 µM of BG95 led to a highly significant shortening of mean mitochondrial length. The length of mitochondria was assessed using the FilamentDetector plugin of ImageJ. Mean mitochondrial length was measured in at least 5 individual images and >100 cells per compound/concentration. Data are given as the mean of mitochondrial length + SD. Statistical analysis was performed using ordinary One-way ANOVA followed by post-hoc Tukey’s test compared to untreated. **** p < 0.0001; *** p < 0.001; n.s., not significant (D–F) Representative confocal images of mitochondrial structure (green) in HFFs treated with GCV, ART or BG95, respectively (all 10 µM). Note in (F) the BG95 autofluorescence (blue) signal colocalized with mitochondria (green).