Fig. 6

MKPC conditioned medium promote angiogenic process in vitro. a, b Conditioned medium from MKPCs ameliorates endothelial cell death following hypoxic culture condition. Endothelial cells (MMECs) were treated with or without conditioned medium from MKPCs (MKPC CM) under normoxic or hypoxic culture conditions for 24 or 28 hours. Cell death was measured by Annexin V and PI staining detected by flow cytometry. Each experiment was repeated three times. Cells in the upper left and upper right plots were considered death cells. c–f Conditioned medium from MKPCs prevents TGF-β-induced EndoMT in vitro. Confocal microscopy demonstrates DAPI (blue), CD31 (green), and α-SMA (red) in MMECs cultured with or without MKPC CM and with or without TGF-β for 14 days c. Representative western blot and relative bar graph analysis of CD31, α-SMA d, e, Smad3 and pSmad3 f protein level in MMECs after various treatments. Fibroblast-like change of MMECs was observed following TGF-β treatment, but the change was reduced in MMECs incubated with MKPC CM c. TGF-β treatment (5 ng/ml) caused increased α-SMA protein expression and decreased CD31 protein expression d, e. MKPC CM inhibited phosphorylation of Smad3 in MMEC after 1 hour of TGF-β stimulation f and attenuated these TGF-β-induced EndoMT responses. *P <0.05 compared with control or between two groups. α-SMA alpha-smooth muscle actin, DAPI 4′,6-diamidino-2-phenylindole, MMEC mouse pancreatic microvascular endothelial cell, TGF-β transforming growth factor beta