FIG 7.

Characterization of NPC1⁺ endosomes in BSC-1 cells. BSC-1 cells were fixed, permeabilized, and stained with a rabbit monoclonal antibody against NPC1 as well as mouse monoclonal antibodies versus CD63, Lamp1, LBPA, M6PR, or EEA1, as indicated. They were then stained with corresponding Alexa Fluor 647 anti-rabbit or anti-mouse Alexa Fluor 546 secondary antibodies, as appropriate, and observed by confocal microscopy. For each sample, 10 random fields were analyzed for overlap between NPC1 and the indicated marker. (A, C) Manders colocalization coefficients. Error bars represent SD. Representative images (B, D) are shown below each graph. The data are from two separate experiments shown on the left (A, B) and right (C, D). In another experiment, high colocalization was also seen between Lamp2 and NPC1 (not shown).