Figure 2
From: Identification of the angiogenic gene signature induced by EGF and hypoxia in colorectal cancer
Angiogenic gene expression in Caco-2 cells exposed to hypoxia is HIF-1 dependent. Caco-2 cells were transfected with siRNA targeting luciferase (siLuc), HIF-1a (siHIF-1a) or HIF-2a (siHIF-2a), and subsequently exposed to hypoxia (1% O2) for 24 hours. Changes in (a) ANGPTL4, (b) EFNA3, (c) TGFβ1 and (d) VEGF mRNA levels were determined by Q-PCR using the 2-ΔΔCt method and are expressed relative to HKG 18S. Data are mean ± SEM from 3 representative experiments, and were analysed by 1-way ANOVA of ΔCt values versus siLuc: ns = not significant, *** p < 0.001. Dashed line shows response of cells exposed to normoxia (21% O2). (e) Western blots, demonstrating specific HIF knockdown at the protein level, with α-tubulin shown as loading control. N = normoxia, U = untransfected cells exposed to hypoxia.