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. 2012 May 30;109(24):9611–9616. doi: 10.1073/pnas.1112368109

Fig. 4.

Fig. 4.

Overexpression of WT ATP13A2 in ATP13A2-deficient cells rescues lysosomal deficiency. (A) Overexpression of ATP13A2 assessed by Western blot in stable cell lines. (B) Colocalization of LysoTracker staining and GFP immunofluorescence in sh403-1 cells transfected with ATP13A2-GFP. (Scale bars, 10 μm.) (C and D) Cath-D activity in lysosomal fractions from sh403-1 cells and mutant ATP13A2 fibroblasts overexpressing or not overexpressing WT ATP13A2. Effects of ATP13A2 overexpression on lysosomal stress susceptibility to lysosomal inhibitor NH4Cl (40 mM for 24 h) in sh403-1 cells and mutant ATP13A2 fibroblasts (L3292), as evidenced by the presence of acid phosphatase (E and G) and β-hexosaminidase (F and H) activities in the cytosol of NH4Cl-treated cells after removing lysosomes by differential centrifugation. (I) Cell death in sh403-1 cells overexpressing or not overexpressing ATP13A2. In all panels, n = 3–5 per experimental group. *P < 0.05 compared with control (Ctrl) untreated (UT) cells; #P < 0.05 compared with control treated cells.