Immunocytochemistry of (A.) type II cells or (B.) alveolar macrophages isolated from wild type and NPC mutant mice.

<p>A. Type II cells were isolated from NPC1 (Top) or NPC2 (Bottom) mutant mice or their corresponding wild type littermates and placed in culture for 24 hrs. The cells were fixed and stained with anti-ABCA3 antibody (green) to mark the lamellar body limiting membrane or filipin (Fil, gray or blue) to mark cholesterol. The merged pictures with anti-ABCA3 in green and filipin in blue are enlarged. Scale bar = 5 µm. <b>B</b>. Alveolar macrophages from NPC mutant mice contain cholesterol. Alveolar macrophages were isolated from the lung lavage from NPC1 (Top) or NPC2 (Bottom) mutant mice or their corresponding wild type littermates and placed in culture for 2 hrs. The cells were fixed and stained with filipin which labels free unesterified cholesterol. Lt, Phase micrograph; Fil, Filipin stain in gray. Merge of phase and filipin (blue) are enlarged. Scale bar = 10 µm.