Fig. 4

Effect of UFH pretreatment on LPS-induced disassembly of MTs. HPMECs were pretreated with vehicle or UFH (10 U/ml, 30 min) followed by LPS (10 μg/ml) stimulation for 6 h. a MT remodeling was examined by immunofluorescence staining with Alexa Fluor® 488, (× 60, scale bars = 10 μm). LPS stimulation triggered striking MT disassembly, which was prevented by UFH pretreatment. b Western blotting was used to detect polymeric/monomeric tubulin ratio. GAPDH was used as the internal control for monomeric and polymeric tubulin. UFH pretreatment decreased monomeric tubulin content and increased tubulin polymerization. c Acetylated tubulin was examined by immunofluorescence staining with cyanine 2, (× 60, scale bars = 10 μm). UFH increased the tubulin acetylation in HPMECs treated with LPS. d Pool of stable MTs was determined by western blot analysis with antibodies against acetylated tubulin. UFH pretreatment inhibited the LPS-induced decrease in acetylated tubulin level. Data are representative of three independent experiments and are shown as mean ± SD. t-test, * P < 0.05 compared to the vehicle group. t-test, # P < 0.05 compared to the LPS-treated group