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. 2015 Nov 23;5:17035. doi: 10.1038/srep17035

Figure 3. Ability of LNP to silence target gene in the mouse liver.

Figure 3

LNP (with Toc-siRNA including Toc-siApoB targeting to mouse ApoB mRNA, Toc-siOAT3 targeting to mouse Oat3 mRNA, or Toc-siScr scrambled sequence of Toc-siApoB) or MM (without Toc-siRNA) was rectally administered as enema to the mice at a dose of 10 mg/kg of Toc-siRNA three times with 2-h intervals. (a) Quantitative RT-PCR analyses of endogenous ApoB, Gapdh, and Hprt mRNA levels were performed 24 h after final administration. (b) Dose-dependent reduction of ApoB mRNA levels in the liver after administration of different doses of LNP. The ApoB mRNA levels (normalized to Gapdh mRNA levels) were determined 24 h after final administration. (c) Quantitative RT-PCR analysis of liver ApoB mRNA levels relative to Gapdh mRNA performed at the indicated time points after final administration. (d,e) Western blot analysis to detect ApoB100 and ApoB48 (d), and LDL-cholesterol and triglyceride levels in serum (e) 24 h after final administration. All experiments were independently performed in duplicate. The quantitative data were normalized to administration of mock mixed micelles alone (MM) and shown as the mean values ± s.e.m. (n = 3). *P < 0.05. (f) Agarose gel of 5′-RACE–PCR amplification, showing specific cleavage products in the liver.