Nano-Scaffold for Therapeutic Angiogenesis Figure 6. Expression of proangiogenic factors in ischemic hind limb muscles treated with transplantation. Intramuscular levels of proangiogenic factors in whole thigh muscles were quantified in hind limb ischemic model (BALB/cAJcl mice) 3 and 7 days after ischemic induction. Vehicle alone, NS alone, BMNCs alone, LA+BMNCs or NS+BMNCs were injected into ischemic thigh muscles simultaneously with ischemic induction. Intramuscular levels of proangiogenic factors were corrected for total protein and expressed in arbitrary units (n = 6 in each group). Data are shown as means (SD). *, P,0.05 for the NS+BMNCs group compared to the non-ischemic muscle (sham), vehicle, NS, BMNCs and LA+BMNCs groups. **, P,0.05 for the NS+BMNCs group compared to the non-ischemic muscle (sham), vehicle, NS and BMNCs groups. ***, P,0.05 for the NS+BMNCs group compared to the non-ischemic muscle (sham), vehicle and NS groups. Abbreviations: NS, nano-scaffolds; LA, unmodified PLLA microspheres; BMNCs, bone marrow mononuclear cells. doi:10.1371/journal.pone.0035199.g006 Figure 7. Colocalization of transplanted BMNCs and VEGF (A) or FGF-2 (B). Tissue sections from hind limb ischemic mice (C57BL/6NCrSlc) 7 days after transplantation of NS+BMNCs were immunofluorescently stained using anti-mouse VEGF antibody (A) or anti-mouse FGF-2 antibody (B) and DAPI. BMNCs were derived from EGFP-transgenic mice. Scale bars, 100 mm. Abbreviations: NS, nano-scaffolds; LA, unmodified PLLA microspheres; BMNCs, bone marrow mononuclear cells. doi:10.1371/journal.pone.0035199.g007 PLoS ONE | www.plosone.org 8 April 2012 | Volume 7 | Issue 4 | e35199