Figure 1.

Assessment of EV release and lysosomal alterations in fibroblast cell cultures. (A,B) EVs were isolated by sequential centrifugation from surnatant of fibroblasts from non-mutated (NM-PD) and GBA-mutated (GBA-PD) PD patients, carrying mild (N370S) or severe (L444P) mutations, and healthy subjects (HC). Nanoparticle tracking analysis was used to determine the concentration of (A) medium and (B) small EV released. Data were expressed as number of vesicles secreted per cell. (C–E) Evaluation of (C) glucocerebrosidase and (D) cathepsin D (CathD) activity by fluorimetric assay, and (E) Western blot analysis of pro-CathD and mature CathD (mCathD) levels. The ratio between pro/mCathD was also reported in the bar graph. All data were expressed as mean ± SEM of three independent experiments (n = 2 for cathepsin D turnover).