Fig 2: Fibroblasts from SPG11 patients also exhibit enlarged LAMP1-positive structures. (A and B) Confocal microscopy analysis of the fibroblast cell lines stained with LAMP1 antibodies: (A) control and SPG15 fibroblasts; (B) SPG11 fibroblasts. Insets in the left panels are enlarged at the right. Nuclei are stained with DAPI. (C) Quantification of lysosome circumference from subject-derived cells, mean ± SEM (n = 3 independent experiments, each evaluating at least 250 lysosomes). (D) EM of SPG11 fibroblasts. Scale bar: 500 nm. (E–G) LC3-positive autophagosomes were counted in the indicated fibroblast lines (mean ± SD, n = 3 independent experiments, each evaluating at least 200 cells). DAPI staining identifies the nucleus. Scale bar: 10 µm. ***P < 0.001; **P < 0.01; *P < 0.05; paired Student's t test.

Fig 3: Both spastizin and spatacsin proteins are depleted from SPG15- and SPG11-patient cell lines. Cell extracts prepared from SPG15 (II-2) and SPG11 patient cell lines were immunoblotted (IB) as shown. PLC?1 and ß-tubulin levels were monitored as controls for protein loading. Asterisks (*) identify cross-reacting proteins. Migrations of protein standards (in kDa) are at the left.