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Figure 1 | BMC Cancer

Figure 1

From: HIF2α reduces growth rate but promotes angiogenesis in a mouse model of neuroblastoma

Figure 1

Characterization of native and transfected N1E-115 cells. a) Expression of HIF1α, HIF2α, VEGF, Neuropilin-1 (NRP-1), adrenomedullin (ADM), neuronal-specific enolase (NSE), Id2 and Chromogranin A (CgA) as well as HIF2α endothelial-specific targets VEGFR-1, VEGFR-2 and Tie2 in N1E-115 cells and mouse kidney cDNAs. DNA ladder (L) is indicated in base pairs. b) Comparison of the human HIF2α (hHIF2α) 870 amino acids protein with the HIF2α(1–485) mutant shows that both activation domains have been truncated in the dominant negative form. c) Specific RT-PCR amplification of hHIF2α sequence was used to confirm the efficiency of transfection with both hHIF2α (E1 and E2) and hHIF2α(1–485) (DN5 and DN8) expressing vectors. Western blot using a specific anti-human HIF2α antibody confirmed the nuclear presence of the wild-type protein in E1 and E2 clones. d) Tyrosine hydroxylase (TH) expression was assessed by western blot and e) VEGF protein expression quantified by ELISA in duplicate 21% or 4% O2 atmosphere. Asterisks on bars illustrate statistical significance compared to NT cells in the same oxygen condition. Upper asterisks indicate significance between 21% and 4% O2 for each clone. f) Western blot evaluation of HIF1α levels revealed no variation between the different cell clones in both 21% and 4% O2.

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