Figure 5. Combination treatment of PEG-lipid and HPBCD lead to synergistic clearance in Npc1^−/− cells.

Npc1^−/− MEFs cells were treated with DSPE-PEG_2k micelles (0–500 μM) and HPβCD (0–10 mM). Cells were fixed and stained with filipin and nuclear mask. (A) Representative images of select treatments are shown and illustrate the synergistic cholesterol reduction of the two treatments. (B) Cells were quantitatively analyzed for filipin fluorescence per cell, and the dose dependent reduction of filipin fluorescence is shown normalized to untreated cells. Images per well were taken (n = 3, 16 images/well) and cells were quantified for total filipin fluorescence. The treatment with 500 μM DSPE-PEG_2k and 10 mM HPβCD is toxic, and no cells are observed. Results were tested for statistical significance using one-way ANOVA with Tukey’s multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001 and NS = not significant.