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. 2017 Dec 26;22(3):1583–1600. doi: 10.1111/jcmm.13434

Figure 3.

Figure 3

mTOR pathway regulates chemotaxis and capillary tube formation capacity of BM‐EPCs. (A) Gene expression of VEGF, KDR, PECAM‐1 and VE‐cadherin in Mtp‐treated BM‐EPCs. Cells were cultivated with 1, 10 and 100 μM Mtp or bFGF (50 ng/ml) for 48 hrs and treated with 100 μM Mtp for 24, 48, 72 and 96 hrs. Gene levels were assessed via qRT‐PCR and normalized to β‐actin; related gene was up‐regulated by Mtp at 48 hrs or less; (B–E) Western blot analysis of p‐mTOR, p‐p70S6K, p‐4EBP1, SQSTM1/P62 and Beclin‐1 in different doses of Mtp‐treated BM‐EPCs for 48 hrs. Mtp evidently increased mTOR pathway proteins and decreased autophagy level; (F, G) cell chemotaxis regulated by the treatments of rapamycin and/or Mtp. BM‐EPCs were treated with 100 nM rapamycin for 2 hrs prior to treatment with Mtp for 48 hrs. The numbers of migrated cells were quantified by performing cell counts of 10 random fields (scale bar: 50 μm); (H, I) in vitro tube formation results of BM‐EPCs treated by rapamycin and/or Mtp (scale bar: 100 μm); the densitometric analysis of all Western blot bands was normalized to the total proteins or GAPDH. n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001 versus the indicated group.