Figure 2.

Transfected naked miRNA is rapidly incorporated into RNA-induced silencing complexes in resting platelets. Platelets at 2 × 10⁸/ml were subject to UV crosslinking at the indicated times after co-incubation with ds miR-223–3p mimic, with 10 μM BCI-137 or equivalent volume of DMSO vehicle as shown, and lysed under non-denaturing conditions. RNA-protein complexes containing the biotin-miRNA mimic were enriched by pulldown with neutravidin beads, and proteins in the whole cell lysate (WCL) and bead pulldown fractions were subject to Western blotting with antibodies to AGO2, and β-actin as loading control. (A) Representative blot. (B) AGO2 pulldown/WCL ratios normalized to un-transfected samples, + s.e.m. *, p < 0.02; **, p < 0.05. n.s., not significant. n = 4.