Figure 3  Transferred NMP leads to increased expression of photoreceptor-specific proteins in the rds +/− retina.
(A–C) cDNA was collected at PI-30, and message levels of photoreceptor genes were analyzed by qRT-PCR. (A) CBA-NMP nanoparticle injection leads to a modest increase in Rom-1 message levels, while IRBP-NMP nanoparticle injection increases expression four- to five-fold over levels in uninjected control eyes. (B,C) CBA-NMP and IRBP-NMP nanoparticle injections lead to increases in rod (B) and cone (C) opsins. (A–C); N = 3 animals per group. (D–F) Protein levels at PI-30 after nanoparticle injection were examined. Representative SDS-PAGE/Western blots from individual retinas are shown (N = 5–6 animals per group). (D) CBA-NMP and IRBP-NMP nanoparticle injections increase RDS and ROM-1 protein levels (protein load: 20 µg per lane). (E) Increases in rhodopsin protein (RHO) are detected after injection of both CBA-NMP and IRBP-NMP nanoparticles (protein load: 10 µg per lane). (F) No change in S-opsin (S-ops) protein level is detected after nanoparticle injection (protein load: 50 µg per lane). (G) Double immunolabeling for transferred RDS (mAB 3B6, green) and cone OSs (S-opsin, red) with nuclear counterstain (DAPI, blue) was performed on frozen sections from PI-30 eyes. Representative cones from two different animals are shown for each treatment. Cones in eyes injected with CBA-NMP or IRBP-NMP nanoparticles express transferred NMP (top and middle rows). Saline injected eyes express no transferred NMP (bottom row). Scale bar, 5 µm; N = 3–5 animals per treatment group. Abbreviations: OS, outer segment layer; IS, inner segment layer; ONL, outer nuclear layer * = p<0.05.