Fig. 7. COMP-Ang1 induces SDF-1/CXCR4/HIF-1α mediated angiogenesis. (A, B) Confocal images of CXCR4 (green) with DAPI for nucleus marker (blue)with/without COMP-Ang1 treatment in EPC before or after FAK or AKT silencing or mTOR inhibition (A) and densitometric analyses (B). Scale bar, 200 µm. All values are the means±SEM. ***p<0.001. (C, E) Confocal images of SDF-1 (red) and FLAG (green) with DAPI of nucleus marker (blue) in ischemia rat with/without EPC treatment in the absence or presence of co-treatment with COMP-Ang1 (C) followed by densitometric analysis (E). Scale bar, 20 µm. All values are the means±SEM. **p <0.01, *p<0.05. (D) The time-dependent changes of extracellular (supernatant) SDF-1 in EPC and HUVEC, treated with COMP-Ang1 for 1, 3, 6, 12, 24 h, were determined by ELISA. All values are the means±SEM. *p<0.05. (F) The relative mRNA expression level of HIF-1α before and after COMP-Ang1 treatment in EPC with/without Tie2, FAK, or AKT silencing or mTOR inhibition. All values are the means±SEM. ***p<0.001. (G) Western blot analyses of HIF-1α protein before or after COMP-Ang1 treatment in EPC with/without Tie2, FAK, or AKT silencing or mTOR inhibition. Whole cell lysates (30 µg protein/lane) were subjected to Western blot analysis to determine the levels of HIF-1α. β-actin was used as an internal control for equal protein loading for each lane. The mRNA and protein expression levels of HIF-1α were increased dramatically after COMP-Ang1 treatment, but that of HIF-1α no changed after Tie2 and AKT silencing.
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