Fig 5. Effect of recombinant enzyme on the ALP pathway and ATP levels.
(A-B) Autophagy in PBMC (A) or lymphocytes (B) following recombinant GCase treatment. PBMCs or lymphocytes from healthy subjects (control, black bar) and GD1 (grey bar) patients were treated for 3h with 10 μg/ml rhGCase and stained with Cyto-ID autophagy detection kit. (C) Autophagosome staining of PBMCs and lymphocytes (from healthy subjects) incubated with or without 10 μg/ml rhGCase. Staining was performed using Cyto-ID autophagy kit (green color). (D and E) Effect of recombinant GCase and α-Gal A on lysosomes. PBMCs from healthy (black bar) and GD1 (grey bar) individuals were incubated for 3h with the 10 μg/ml rhGCase or α-Gal A and stained with lysosomal dye, LysoTracker Red. (F) ATP level in healthy (control) and GD type 1 PBMCs before and after 3h rhGCase treatment. (G) ATP level in control and FD PBMCs before and after 3h α-Gal A treatment. All samples were measured in triplicates, n = 3–4 patients from each group. All data were expressed as S.E.M. and * p<0.05 vs untreated samples.