Figure 5: Sustained in vivo ZNF304 gene silencing.

(a) Size and (b) zeta potential of DANP determined by Zeta Sizer. (c) Atomic force microscopy images of DANP showing the morphology and size distribution of particles. (d) Biodistribution of rhodamine 6G-labelled DANP in vivo. Tumour and the major organs were removed 24 h after a single administration of rhodamine 6G-labelled DANP. The nanoparticles were monitored using fluorescent microscopy and representative images were taken at × 20 (left) and × 40 magnification (centre). Scale bar, 100 μm. Number of nanoparticles was counted at five fields per slide (right). Data are presented as means±standard error of the mean (s.e.m.). (e) Sustained in vivo ZNF304 silencing in HeyA8 orthotopic model of OC. Tumours were removed and analysed by immunoblotting at 3, 7 and 14 days after a single administration of DANP-Control siRNA or DANP-ZNF304 siRNA. (f) Effect of DANP, DANP-Control siRNA and DANP-ZNF304 siRNA on cytokine levels in plasma at 72 h, after a single intravenous administration. Inflammatory cytokine responses were assessed in the serum of C57 black mice. Mice were treated with single i.v. injections of DANP alone (n=6), DANP-Control siRNA (n=6), DANP-ZNF304 siRNA (n=6) and no treatment (n=2), and serum was collected after 72 h using cardiac puncture. A Luminex assay designed to detect 12 pro-inflammatory cytokines was used.