Fig. 5
From: SOD3 improves the tumor response to chemotherapy by stabilizing endothelial HIF-2α
HIF-2α mediates SOD3-induced VEC transcription. a Relative luciferase activity (RLU) in 1G11 cells cotransfected with SOD3 (or mock) and the full-length VEC promoter or deletion mutants. Values were normalized to the signal in 1G11-mock cells for each promoter (n = 4). b Full-length (~2.5 kb) VEC promoter with binding sites for the main transcription factors. Numbers indicate positions relative to the transcription start site. HRE hypoxia response element, SBS Sox-binding site, EBS Ets-1-binding site. c ChIP analysis of VEC promoter in mock- and SOD3-transfected 1G11 cells using anti-HIF-2α and -KLF-4 antibodies. Coprecipitated DNA fragments were PCR-quantified using primers in a (arrows; n = 5). d RLU in 1G11-mock or -SOD3 cells transfected with VEC promoter HRE mutants. The mutated HRE site is indicated (white square; n = 10). e Immunoblot analysis of control (shNT) and HIF-2α-silenced (shHIF-2α) 1G11 cells (n = 3). The HIF-2α:tubulin densitometry ratio is shown (bottom). f VEC mRNA levels, using CD31 as reference, in cells as in e. g Relative VEC promoter activity in control and shHIF-2α-1G11 cells transfected with mock or SOD3 plasmids. Data shown as mean ± SEM (b-g). *p < 0.05, ***p < 0.001; two-tailed Student’s t-test
