Figure 8: GBA-PD and GD iPSC-derived neurons show alterations in the autophagic/lysosomal system.

(a) Immunostaining of differentiated iPSC cultures at DIV65 under basal conditions. Cells were stained for TH (green), β-TubIII (magenta) and LAMP1 (red). Cell nuclei were counterstained with Hoechst (blue). High magnification insets are shown in the right lane. (Bars, 20 μm and 10 μm in the insets.) (b) Average size and number of LAMP1⁺ particles in TH⁺ neurons were quantified by ImageJ. Data are represented as mean+s.e.m.; experiments were independently repeated three times in triplicate. *P<0.01, **P<0.001, one-way ANOVA. (c) Western blot analysis for LC3 in iPSC-derived neuronal cultures at DIV65, untreated (−) or treated with 200 μM leupeptin and 20 mM NH₄Cl for 4 h (+). (d) Quantification of the basal levels of LC3-II relative to β-actin. Data are represented as mean+s.e.m.; experiments were independently repeated three times. *P<0.01; **P<0.001, one-way ANOVA. (e) Quantification of LC3 flux normalized to actin. Data are represented as mean+s.e.m.; experiments were independently repeated three times in triplicate. *P<0.01; **P<0.001, one-way ANOVA.