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. Author manuscript; available in PMC: 2013 Jun 13.
Published in final edited form as: Oncogene. 2012 Feb 6;31(50):5132–5143. doi: 10.1038/onc.2012.16

Figure 1. EphB2 overexpression promotes GBM neurosphere cell migration.

Figure 1

(A). Western blot analysis showed that EphB2 expression in GBM neurosphere lines HSR-GBM1A and GBM1B was elevated compared to that in normal neurospheres from fetal brain. (B). Reverse transcripase-PCR detected expression of EphB2 ligands, ephrin-B1, B2, B3, but not ephrin-A5, in HSR-GBM1A and HSR-GBM1B cells. (C). Immunoblot analysis showed that compared to control transfected cells (Ploc), there was a ~2 fold increase in EphB2 expression in GBM-1A cells transfected with EphB2 cDNA (EphB2-OVE). (D). Phase contrast and fluorescent images of GBM neurospheres transfected with control plasmid ploc and labeled with RFP (upper panels) or with EphB2 cDNA and labeled with GFP (lower panels). Bar=100 μm. (E). Ploc and EphB2-OVE cells were treated with EphB2 ligand ephrin-B1/Fc (1μg/ml) or human Fc (1μg/ml). Protein extracts were immunoprecipitated with anti-EphB2 antibody and EphB2 phosphorylation was detected with anti-phosphoylated-tyrosine antibody. In EphB2-OVE cells, there was a strong activation of EphB2 receptor after ligand binding, indicating functional EphB2 receptors in GBM neurospheres.