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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Gene Ther. 2015 Jul 24;23(1):1–9. doi: 10.1038/gt.2015.81

Figure 5.

Figure 5

Panel a. Significant reduction in cerebral atrophy (measured during the 4th week after BCAO, purple bar) in the scAAV2-CMV-hG-CSF treatment groups with multiple doses (green bar, n = 5 survivors of 5) compared to the placebo group (blue bar, n = 7 survivors of 28) or BCAO+ single dose hG-CSF (red bar, n = 5 survivors of 8). We validated the expression of hG-CSF antigen using immunohistochemistry in the 11th week (Panels c & d). Panel b. A unique ROI (arrow) using SPION-AS-hG-CSF one week after the final scAAV2-CMV-hG-CSF treatment (the 3rd week). Panel b1. The ROI in delta R2* maps from both the multiple treatment (n = 5) and placebo (n = 4 survivors of 16, Panel b2) groups of mice. Panels c & d. hG-CSF expression in the proximity of the ROI identified in vivo (b1), with no expression found in the placebo group (b2 & d). Mice with BCAO only did not show elevated hG-CSF expression (n = 2 survivors of 8, data not shown). Panel e. NeuN expression in small blood vessels (Cy3-GSL I) in mice with AAV2-CMV-hG-CSF (BCAO+hG-CSF, n = 3) bar = 100 µm. NeuN expression (Cy2, arrow) occurs mostly in the neuronal nuclei of control and placebo groups (BCAO, n = 2 survivors of 8; BCAO+GFP, n = 4 survivors of 16). Panel f. T1-weighted MRI of mice (n=4) before and after application of eye drops containing non-targetng MR-CA (Gd-DTPA, 500 mM in 10 µl Magnevist, Scherling, Berlin, Germany) to the conjunctival sac; the distribution of MR-CA is shown in the same mouse. Arrow: the naso-lacrimal canal, #: the vitrous chamber; asterisk: lens. Panel g. The distribution of targeted MR-CA (Gd-nestin) in longitudinal MR acquisition (n = 2). T1-weighted MRI was acquired before (g1), 10 min (g2), 180 min (g3) and 41 hr (g4) after application of Gd-nestin (50 µg Gd-NA and 30 pmol biotin-sODN-nestin per 2 µl per mouse to one eye) in mice with BCAO-60. Arrows: chorid of the retina; * both hemispheres of the brain. Biotinylated sODN-nestin binds to NeutrAvidin (NA) on SPION or Gd and the MR-CA targets Nestin mRNA of percytes of neovessels after BCAO-6014,15.