adck3 mutant cells display an increase in lysosomal content but no effect on lysosomal pH under basal conditions.

(A). Ultrastructural analysis of adck3 mutant cells via TEM. An increase mitochondrial electron density and an increase in vesicular structures indicative of lysosomal expansion is observed in FD130 cells. In left panels, arrow heads indicate vesicular structures. In right panels, arrows indicate the position of electron dense mitochondria and arrow heads indicate electron dense lamellar bodies. (B, C, D). adck3 mutant fibroblasts display a significant increase in lysosomal staining. Cells stained with lysotracker red (B) and acridine orange (C), or processed via β-galactosidase X-Gal staining were analysed via microscopy and flow cytometry. Results in (C) expressed as % acidic vesicle (AVO) positive cells (i.e. high AO-Red, low AO-Green–See grey shaded box in accompanying dotplots) normalised to control values ± S. E. M. n = 3. White bars: 30 μm. Images in (D) at 5x mag. (E). adck3 cells show no change in lysosomal pH compared to control. Cells were stained with Lysosensor blue/yellow dextran for 24 h, chased for 6 h and analysed using a plate reader. n = 3. * p p