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. 2018 Sep 24;28(5):273–284. doi: 10.1089/nat.2018.0722

FIG. 3.

FIG. 3.

LNA ASOs targeting NPC1 reduce expression of NPC1 mRNA in HeLa cells. (a) Six LNA ASOs targeting NPC1 were transfected into HeLa cells and RNA harvested 48 h later to measure mRNA levels. The negative control LNA ASO, siRNAs targeting NPC1, and nontargeting siRNAs were included as controls. LNA20, LNA21, and LNA22 demonstrated greatest reduction in NPC1 mRNA levels. (b) LNA ASOs targeting NPC1 reduce NPC1 protein levels in HeLa cells. One hundred nanomolar of LNA ASOs targeting NPC1 (LNA20, 21, and 22) were transfected into HeLa cells, and protein harvested 48 h later and NPC1 levels measured by immunoblotting. Tubulin was used as loading control. (c) LNA20 targeting NPC1 reduces expression of NPC1 mRNA in mouse macrophages. NPC1 siRNA was used as positive control. The negative control LNA24 and nontargeting siRNA were used as negative controls. (d) LNA20 targeting Npc1 reduces Npc1 protein levels in mouse macrophages as assessed by immunoblotting. β-Tubulin was used as loading control. Npc1 siRNA was used as positive control for Npc1 knockdown. Negative control LNA ASO and nontargeting siRNA controls did not reduce Npc1 levels. ***P < 0.001. Experiments were performed in biological triplicates; each biological replicate was performed in technical triplicates. NPC1, Niemann-Pick C1.