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. 2020 May 13;69(8):1779–1792. doi: 10.2337/db19-0829

Figure 2.

Figure 2

Endothelial MT overexpression preserves EPC mobilization and homing and alleviates oxidative stress in diabetic HLI. T2D was induced as described in Fig. 1. At day 3 after HLI surgery, peripheral blood was collected to evaluate the percentage of EPCs (CD34+/VEGFR2+) in circulation by a flow cytometry assay (A and B); transverse sections of gastrocnemius muscle were stained by CD34 and VEGFR2 (C) to evaluate EPC infiltration (arrow); the infiltration incidence was expressed as CD34+/VEGFR2+ cells per high-power field (HPF) (D). The plasma SDF-1 (E) and VEGF (F) levels were detected by ELISA. The expression of SDF-1 (G and H), VEGF (G and I), and HIF-1α (G and K) and phosphorylation of Akt (G and J) 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 (L), and the fluorescence intensity was quantified by Image J (M). Data are mean ± SD. n ≥ 4 mice/group. *P < 0.05 vs. WT control (Ctrl); #P < 0.05 vs. WT diabetes mellitus (DM).