Fig. 2

TMTC3 was regulated by HIF-1α and modulated the angiogenesis of HUVECs. A. Schematic diagram depicting the positions of the binding sites of HIF-1α in the TMTC3 promoter from the JASPAR database. TSS, transcription start site. B and C. The levels of TMTC3 was increased both in protein (B) and RNA level (C) after transfected with HIF-1α. D. Quantification of ChIP analysis of the interaction between HIF-1α and TMTC3 promoter. IgG was used as a negative control. E. Dual-luciferase reporter assay demonstrating the effects on TMTC3 transcription activity in the indicated cells following HIF-1α overexpression. F. Conditioned medium from TMTC3 knockdown cells decreased HUVECs capillary tube formation. The capillary length and number of branches were counted by Image J software. G and H. ROS accumulation was analyzed by flow cytometry with DCFH-DA probe staining in KYSE450 (G) and KYSE410 (H) cells. The fluorescence intensity and positive cells in each group were calculated. All data are expressed as the mean ± SD. *, p < 0.05; **, p < 0.01; ***, p < 0.001. n = 3