Fig. 5

The role of HIF-1α in LRG1-induced EMT and VEGF-A expression. a HIF-1α mRNA expression in HCT116 and SW480 cells stimulated with indicated concentrations of LRG1 for 24 h was measured by RT-PCR. β-actin was used as the internal control. b The protein levels of HIF-1α, Twist1, E-cadherin, and N-cadherin in response to LRG1 in SW480 cells were measured by western blot. c HIF-1α expression was knocked down in siRNA-transfected SW480 cells at both mRNA and protein levels. d SW480 cells were transfected with HIF-1α or control siRNA for 24 h, and treated with or without rLRG1 (500 ng/ml) for additional 24 h. Secretion of VEGF-A from SW480 cells was quantified by ELISA. e HIF-1α, VEGF-A, E-cadherin, N-cadherin and Twist1 expressions were detected by western blot. f SW480 cells were harvested and applied for transwell invasion assays. The numbers of invasive cells (five random fields) were expressed as means ± SEM of three independent experiments. *Compared to the control group, P < 0.05. Scale bar represents 100 μm