Fig. 4

Effects of deer TMSB10 on cell proliferation, tube formation, and motility of HUVECs as well as on DRG outgrowth. a HUVECs overexpressing deer TMSB10 compared with vector alone. HUVECs overexpressing deer TMSB10 had significantly higher growth rates than that of the vector control group. b Determination of HUVEC proliferation using the MTT assay. HUVEC cells were exposed for 24, 48, and 72 h in the presence of deer exogenous TMSB10 (eTMSB10: 50, 100, or 150 ng/ml) and compared with control. The growth rates of the HUVECs treated with deer eTMSB10 (50, 100, and 150 ng/ml) were significantly higher than in controls. c Representative images (inverted phase contrast) of tube formation assays. Scale bars = 200 μm. d Quantification of tube formation by calculating the average number of branched vessels per field of view in response to exogenous TMSB10 (eTMSB10: 50, 100, or 150 ng/ml), overexpressed TMSB10 (TMSB10), and vascular endothelial growth factor (VEGF) as a positive control. Results showed that deer eTMSB10 and deer TMSB10 overexpressing HUVECs showed more tube formation in the HUVECs compared with controls or vector. e Transwell migration assays with HUVECs treated with deer eTMSB10 (50, 100, and 150 ng/ml) or overexpressing deer TMSB10 were compared with their controls. Results showed that deer eTMSB10 and deer TMSB10 overexpressing HUVECs showed more migration compared with the control or vector. f Lamellipodium emerging from a DRG neuron after treatment for 2 days or 5 days with 50, 100, and 150 ng/ml eTMSB10 or nerve growth factor (NGF; 50 ng/ml) as a positive control. Scale bar = 100 μm. g Quantification of average length of lamellipodium. Results showed that deer eTMSB10 significantly increased the growth of lamellipodium from DRG compared with the control. Data represent mean ± SD of three or four experiments. *P < 0.05, **P < 0.01, compared with control or vector (control or vector control = 100%). OD optical density