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Fig. 5 | BMC Cancer

Fig. 5

From: Ascorbic acid and ascorbate-2-phosphate decrease HIF activity and malignant properties of human melanoma cells

Fig. 5

Effect of ascorbic acid on EDHB induced HIF-1 transcriptional activity in melanoma cells. WM9 metastatic melanoma cells were transiently transfected with an HIF-1 HRE-luciferase reporter vector. a Transfected cells were treated for 24 h with 0.5, 0.75 and 1.0 mM EDHB. Induction of HIF-1 transcriptional activity was measured by luciferase assay. EDHB at 0.75 mM was found to be the lowest dose capable of generating near-maximal induction of HIF-1 transcriptional activity and was thus chosen for subsequent experiments. b Cells were treated for 24 h with 0.75 mM EDHB alone (EDHB), 100 μM AA alone (AA), pretreated with 100 μM AA for 4 h prior to addition of EDHB (AA-EDHB), or treated with100 μM AA and EDHB concurrently (EDHB/AA). AA effectively reduces EDHB induced HIF-1 transcriptional activity; with AA pretreatment showing increased efficacy at inhibiting EDHB induced HIF-1 activity vs. concomitant treatment. All HRE-luciferase activity was normalized to β-galactosidase activity. Data are represented as mean ± SEM of n = 3, analyzed by One-way ANOVA followed by Tukey’s multiple comparisons test; * denotes significant difference from control, p < 0.0001, # denotes significant difference from EDHB treatment alone, p < 0.0001

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