Figure 6.

Lysosome structural alteration in MPS VII NPCs and neurons. (A) Immunofluorescence microscopy of control and MPS VII NPC, stained for LAMP1 (green) and DAPI (blue; left panel) or phalloidin (red) and DAPI (blue; right panel); scale bars 20 µm. (B) Immunofluorescence confocal microscopy of control and MPS VII neurons at 1, 3, 6 and 9 weeks of culture, stained for LAMP1 (green), βIII-tubulin (TUJ1, red) and DAPI (blue); scale bars 5 µm. (C) Quantification of acidic vesicles labeled with Lysotracker^TM of control and MPS VII NPC, with areas between 0.1–1.2 μm² per cell (top) and higher than 1.2 μm² per cell (bottom). Mean LAMP1⁺ vesicles content (D), area (µm²) (E) and size distribution (F) per neuron for control and MPS VII neurons along culture time. (G) Immunofluorescence confocal microscopy of control, MPS VII neurons and β-gluc-treated MPS VII neurons at 9 weeks of culture, stained for LAMP1 (green), βIII-tubulin (TUJ1, red) and DAPI (blue); scale bars 5 µm. (H) Mean LAMP1⁺ vesicles size distribution for control, MPS VII neurons and β-gluc-treated MPS VII neurons at 9 weeks of culture.