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A, Control human fibroblasts incubated with LDL for 24 h, immunostained with antipeptide antibodies to the C-terminus of NPC1 protein, and viewed with confocal fluorescence microscopy. NPC1 immunofluorescence is present in small granules that are distributed throughout the cytoplasm of cells. B and C, Control fibroblast incubated with LDL for 24 h and immunostained with both (B) antibodies to a lysosomal membrane glycoprotein (lamp1) and (C) antipeptide antibodies to the C-terminus of NPC1 protein. The colocalization shows that NPC1 protein (C) is present in lamp1-positive vesicles (B). Note that there are many lamp1-positive lysosomes that do not contain NPC1 protein. D and E, Control fibroblast incubated with LDL and U18666A for 24 h. U18666A causes an accumulation of unesterified cholesterol in lysosomes and Golgi of control cells similar to the cholesterol lipidosis in NP-C cells. Cells Source: Niemann-Pick Disease Type C: A Lipid Trafficking Disorder, The Online Metabolic and Molecular Bases of Inherited Disease were stained with both filipin (D), to visualize unesterified cholesterol, and immunostained with (E) antipeptide antibodies to the C-terminus of NPC1 Valle D,NPC1 Beaudet AL, (E Vogelstein B, isKinzler KW, Antonarakis SE, Ballabio A, Gibson The Online Molecular protein. AfterCitation: drug treatment protein at arrows) present in cholesterol-containing lysosomes (DK, atMitchell arrows).G. Bar = 10 μm. Metabolic See Color and Plate 7. Bases of Inherited Disease; 2014 Available at: http://mhmedical.com/ Accessed: August 03, 2017 Copyright © 2017 McGraw-Hill Education. All rights reserved