Fig. 8.

Impact of Vorinostat on expression and trafficking of human LIPA and NPC1 protein from LIPA mutant fibroblasts. A, Western blotting of LIPA protein from WT and LIPA mutant fibroblasts. Cell lysates were extracted, and equal amount of cell lysates were subjected to SDS-PAGE and Western blotting with mouse anti-LIPA antibody. GADPH was used as a loading control. Graph represents the ratio of LIPA protein (LIPA*/LIPA), ***p < 0.001. B, Western blotting of LIPA protein from DMSO or Vorinostat treated LIPA fibroblasts. LIPA fibroblast was treated with DMSO or Vorinostat (10 μm) for 72 h., cell lysates were obtained and subjected to SDS-PAGE and Western blotting. GADPH was used as a loading control. Graph represents the ratio of LIPA protein (LIPA*/LIPA), n.s (no significant). C, Endo H digestion of cell lysates derived from DMSO-treated WT fibroblasts and DMSO or Vorinostat treated LIPA fibroblasts. Samples were separated in 10% SDS-PAGE and Western blotting with LIPA antibody. D, Endo H digestion of cell lysates derived from DMSO-treated WT fibroblasts and DMSO or Vorinostat treated LIPA fibroblasts. Samples were separated in 10% SDS-PAGE and Western blotting with NPC1 antibody. For equal loading control, GADPH or Hsp90 was used in the Western blotting. Bar graphs: endo H sensitive (white) or endo H resistant (black) glycoforms are quantified as percent of total NPC1 in each lane.