Fig. 3

(A) Live 3?dpf npc2^+/+ and npc2^m/m larvae were stained with LysoTracker Red (LysoR) for 1?h and destained in egg water for 2?h before images were taken. npc2^m/m larva showed intense LysoTracker Red staining of the olfactory placodes (arrowheads). A total of 47 npc2^+/+ and 59 npc2^m/m larvae were examined in three experiments. All examined larvae displayed the same staining patterns as shown here. Scale bars: 100??m (individual); 400??m (overlay). (B) Live double staining of npc2^+/+ and npc2^m/m larvae with YO-PRO-1 (neuromast cell marker) and LysoTracker Red. npc2^m/m larva exhibited intense LysoTracker Red staining along lateral line neuromasts but the overall pattern of neuromast organization was unaffected. A total of 67 npc2^+/+ and 75 npc2m/m larvae were examined from three experiments. All larvae displayed the same staining. Scale bars: 1?mm. (C) Filipin-positive puncta were present in 5 and 7?dpf npc2^m/m larvae, indicating unesterified cholesterol accumulation. A total of 26 npc2^+/+ and 19 npc2^m/m 5?dpf larvae and 23 npc2^+/+ and 28 npc2^m/m 7?dpf larvae were examined in three experiments. All examined larvae displayed the same staining patterns. Scale bars: 200??m.