Figure 8. Intracellular processing mechanisms of Toc-HDO.

(a) Schematic illustration of the construction of 13-mer DNA/31-mer cRNA Toc-HDO. Arrows: presumed cleavage sites of the 31-mer cRNA. (b) Northern blot analysis of 31-mer cRNA sequences of Toc-HDO in livers from 6 mg kg⁻¹ of Toc-HDO or RNase-resistant Toc-HDO (that is, Toc-HDO cRNA molecules in which all of the RNA was replaced by 2′-O-methyl RNA (2′-OMe cRNA))-injected mice. cRNA, Toc-HDO with unmodified cRNA; 2′-OMe, Toc-HDO with 2′-OMe cRNA. (c) qRT–PCR analyses of ApoB mRNA levels in liver injected with 0.75 mg kg⁻¹ Toc-HDO, Toc-HDO with 2′-OMe cRNA or PBS alone. cRNA, Toc-HDO with unmodified cRNA; 2′-OMe, Toc-HDO with 2′-OMe cRNA (n=3, mean values±s.e.m., *P<0.05, **P<0.01 versus PBS). Data are representative of at least three independent experiments each (c). P values were calculated from Student's two-tailed t-test (c).