FIGURE 5.

Evaluation of amine-induced vacuole size in fibroblasts with mutations in NPC1 and NPC2. Indicated fibroblasts were incubated with 70 μm NR for 6 h and visualized by phase contrast microscopy. To the right of each phase image is the computer software-generated threshold image of the amine-induced vacuoles that was used to estimate their average diameter. A, wild type cells are shown. B, cells with mutations in NPC1 are shown. C and D, cells with mutations in NPC2 (both GM17910 and GM18455 cell lines) are shown. E, bars represent the average diameter of NR-containing vacuoles in the indicated cell type. Indicated cells (nocodazole (+NOC), black bars) were treated with the microtubule depolymerizing agent nocodazole (1 μm, 1 h before and concurrent with the administration of NR). Bars represent the average ± S.E. of six different cells obtained in three independent experiments (*, p < 0.05 by unpaired t test).