Figure 5.

Targeting of Tmem97 by RNA interference in Npc1^{tm(I1061T)Dso} mice fails to increase hepatic NPC1 levels or to improve lipid abnormalities. (A & B) Npc1^{tm(I1061T)Dso}mice received intraperitoneal injections of locked nucleic acid antisense oligonucleotides (LNA-ASO) targeting murine Tmem97 or GFP starting at postnatal day 28. Vehicle treated mice served as a second control cohort. Livers were harvested on postnatal day 56. Graphs show mRNA levels of Tmem97 and Npc1 relative to untreated control. Tmem97 mRNA was significantly reduced in Tmem97 LNA-ASO treated mice relative to GFP LNA-ASO treated controls and vehicle treated control. No significant changes in Npc1 mRNA levels were observed (n =  8 Vehicle, 8 GFP LNA-ASO, 7 Tmem97 LNA-ASO, ***P < 0.001). (C) Npc1 protein levels in whole liver lysates harvested at postnatal day 56 from Npc1^{tm(I1061T)Dso}mice treated with LNA-ASO targeting murine Tmem97 (n = 7, T1-7) or GFP (n =  8, GFP1-8), or vehicle (n = 8 Vehicle, Veh1-8) as described in (A). (D) Quantification of lipids from liver lysates (C) using tandem mass spectrometry. Graph shows the percent change in Npc1^{tm(I1061T)Dso}mice treated with Tmem97 LNA-ASO (n = 7) relative to those receiving GFP LNA-ASO (n = 8). None of the changes reached statistical significance.