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. 2010 May;51(5):1023–1034. doi: 10.1194/jlr.M002345

Fig. 3.

Fig. 3.

Levels of cholesterol transported to the inner mitochondrial membrane are unchanged by siRNA-mediated depletion of NPC1. A: NPC1 protein levels in F2-CHO cells transfected with nontargeting siRNA (siNT) or siRNA directed against NPC1 (siNPC1), analyzed by immunoblotting. Tubulin was used as a loading control. Shown is one representative blot of five independent experiments. B, C: F2-CHO cells transfected with nontargeting siRNA (siNT; B) or siRNA directed against NPC1 (siNPC1; C) were stained with filipin to visualize cholesterol accumulation. Bars = 40 µm. D: Pregnenolone formed during 24 h from endogenous cholesterol sources in intact, adherent F2-CHO cells grown in the presence of serum and transfected with nontargeting siRNA (siNT) or siRNA directed against NPC1 (siNPC1) was measured as ng pregnenolone per mg cell protein and is depicted as percentage of the average of siNT. Mean ± SEM, five independent experiments [total n = 13 (siNT) or 16 (siNPC1)].