Fig. 8.

NPC1 and NPC2 KO HeLa cells were plated in 96-well plates, transfected with the indicated siRNA for 72 h, and treated or not with 0.7 mM HPCD for the last 36 h. Cells were fixed, stained with filipin, as well as anti-LAMPl Abs and PI (used for segmentation; not shown), and analyzed by automated microscopy. A: Representative images are shown for filipin. B: The filipin intensity in A was quantified in a lampl mask, and the average filipin intensity per cell was calculated. Data are expressed as the ratio between HPCD-treated and untreated (UT) cells for each condition. Means represent three independent experiments. * P < 0.05; ** P < 0.005.