Figure 4.

MT-overexpressing BM-MNCs involved in ischemic angiogenesis in db/db diabetic mice with HLI. BM-MNCs transplantation was performed as described in Fig. 3. At day 7 after HLI surgery, transverse sections of gastrocnemius muscle were stained by CD31 and MT (A) to evaluate the incorporation of MT-overexpressing BM-MNCs in capillaries (arrows), the capillary density was expressed as CD31⁺ cells per high-power field (HPF), and the incorporation incidence was expressed as CD31⁺/MT⁺ cells per HPF (B). The plasma SDF-1 (C) and VEGF (D) levels were detected by ELISA. The expression of SDF-1 (E and F), VEGF (E and G), and HIF-1α (E and I) and phosphorylation of Akt (E and H) in gastrocnemius muscle were detected by Western blot, with β-actin used as loading control. The transverse sections of gastrocnemius muscle were also used to probe superoxide production by DHE staining (J), and the fluorescence intensity was quantified by Image J (K). Data are mean ± SD. n = 5 mice/group. *P < 0.05 vs. vehicle; #P < 0.05 vs. WT.