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. 2014 Jul 3;9(7):e101749. doi: 10.1371/journal.pone.0101749

Figure 4. Effect of individual Argonautes in vivo.

Figure 4

(A) C57BL/6 mice were i.v.-injected with LNP-formulated siRNA targeting Ago2–3′UTR or with control Luciferase siRNA at 0.5 mg/kg. Animals were sacrificed at indicated time points and Ago2 mRNA was quantified by bDNA assay (mean ± s.d., n = 5). (B) bDNA measurement of mRNA knockdown of Rab5c (by siRNA targeting 3′UTR-near at 0.5 mg/kg) and FVII (by siRNA targeting CDS at 0.2 mg/kg) in wild-type mice (mean ± s.d., n = 4–5) following 4 days of Ago2 knockdown (by siRNA targeting 3′UTR at 0.5 mg/kg). (C) Animals were i.v.-injected with Ago1, Ago2, or Luc siRNA on day 1, Ago1 treated animals were further i.v.-injected with Ago2 on day 2; and Rab5c-3′UTR-near siRNA was introduced on day 5 (where indicated; each siRNA was dosed at 0.5 mg/kg). All animals were sacrificed on day 6. Liver mRNA levels for Ago1, Ago2, and Rab5c were measured by bDNA assay (mean ± s.d., n = 4–5). (D) Mice were i.v.-injected with LNP containing 0.5 mg/kg siRNA against each of Ago1 and Ago2 (or 1 mg/kg of Luciferase control siRNA) twice at week long intervals, at the end of the second week mice were i.v.-injected with Fads1–3′UTR or Luciferase control siRNAs at 0.4 mg/kg. Mice were sacrificed for liver RNA extraction the day after the last injection (mean ± s.d., n = 3).