Fig. 2

Hypoxia induces the production of ROS, which reduces the proliferation, survival rate, migration, barrier integrity and microtubular depolymerization in HUVECs. HUVECs were cultured under normoxia or hypoxia condition with or without pretreatment of N-acetyl cysteine (NAC, 1 mM) or wortmannin (1 μM) for 30 min. a ROS production in HUVECs. b Microscopic observation and BrdU staining assessed the HUVECs proliferation. Bar = 50 μm. c The survival rate (%) of HUVECs determined using the MTT assay. d Permeability of HUVECs was tested in cell culture inserts through determining the fluorescence leaked into lower chamber. All the data were normalized to control group. e Cell migration was tested by transwell method. f HUVECs were processed for TEM to examine microtubule structure. Bar = 0.5 μm. g HUVECs were cultured under normoxia, hypoxia or H₂O₂ conditions with or without pretreatment of 1 mM NAC or 1 μM wortmannin for 30 min, total cell lysates were immunoblotted with antibodies to P-Akt, Akt and P85. Representative results from three independent experiments are depicted here. The relative intensity of band was normalized to P85. The data were presented as the mean ± SEM. *P < 0.05