FIGURE 2.
Silencing efficacy and nonspecific immune stimulatory effects of naked siRNAs and SAMiRNAs. A and B, NIH3T3 cells were treated with AR and CTGF naked siRNAs and SAMiRNAs, and then the expression of AR and CTGF mRNA was measured by real-time RT-PCR and compared with scrambled (Sc) siRNAs or SAMiRNAs, respectively. C, the expression of nonspecific innate immune cytokine was evaluated using mouse PBMCs stimulated by naked siRNAs or SAMiRNAs of AR (siAR), CTGF (siCTGF), β-galactosidase (si-β-gal), and their control siRNA (si-Cont) or SAMiRNA (SAMiRNA-Cont). 5 × 105 mouse PBMCs were seeded onto a 48-well plate and treated with PBS, 5 μm naked siRNAs, 1 or 5 μm lyophilized SAMiRNAs, 1 μm of encapsulated liposomes (SNALPs), Lipofectamine RNAimax only, and concanavalin A (ConA, 20 μg/ml). After 24-h incubation, cell culture supernatants were harvested, and the levels of representative innate immune cytokines (TNF-α, MCP-1, IFN-γ, IL-12(p70), and IL-6) were measured by luminex multiplex screening assay. Data represent mean ± S.E. of a minimum of three separate evaluations with duplicates.