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Fig. 6 | Journal of Biomedical Science

Fig. 6

From: A novel HIF1α-STIL-FOXM1 axis regulates tumor metastasis

Fig. 6

STIL expression is induced by HIF1α under hypoxia. a STIL protein (upper panel) and mRNA levels (lower panel) were analyzed by Western blotting and qPCR method, respectively, in the indicated cells under normoxic (20% O2) or hypoxic (1% O2) conditions, or in HIF1α-knockdown cells under hypoxia. b STIL protein (upper panel) and mRNA levels (lower panel) were analyzed in cells transiently transfected with the indicated constructs under normoxia. c STIL promoter activity was measured by reporter assay in HEK293T cells transiently transfected with the pGL3-STIL promoter-luciferase plasmid and the indicated constructs under the normoxic condition. d STIL promoter activity was measured by reporter assay in CL1-0 cells transiently transfected with pGL3-STIL promoter-driven luciferase constructs encoding wild-type HIF1α, HIF1α DNA-binding site mutants, and/or HA-HIF1α (ΔODD) under hypoxia. e The binding of HIF1α to the STIL promoter was analyzed by the ChIP-qPCR assay in CL1-5 cells under hypoxia. VEGF promoter was served as the positive control for HIF1α binding, and the region of 6.0 kb upstream of STIL transcriptional start site within STIL promoter was used as the unrelated control. Methylation of histone H3 (H3-K9Me3) on SAT2 gene was used as a positive control and IgG as a negative control for the ChIP-qPCR assay. f Clinical correlations between STIL, HIF1α, and SLUG in 200 lung cancer specimens were analyzed by IHC and the data were assessed using Pearson’s correlation. Scale bar: 50 μm. g Overall survival of 1927 lung cancer patients stratified by STIL and SLUG expression was determined by Kaplan–Meier analysis. The median survival of the four groups is shown. Significance is determined by the log-rank test (p < 0.001). Data information: Statistical data in ae represent the mean ± SD (n = 3 independent experiments). Significance is determined by t-test (NS, not significant; **p < 0.01; ***p < 0.001)

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