Fig. 1. Expression levels of EphA3 and ephrin-A on various melanoma cell lines and 293T cells. (A) LK63 human B-cell leukaemia cells, melanoma cells and 293T cells as indicated were analysed for cell-surface expression of EphA3 (a-c,e,g,h) with IIIA4 anti-EphA3 monoclonal antibody (solid lines) or an IgG1 isotype matched nonrelevant control antibody (broken lines), and of ephrin-A5 (d,f) with an EphA3-human Fc fusion protein (solid lines) or with EphA1-human Fc fusion protein (broken lines). Analysed cell populations were gated to exclude propidium iodide (PI)-positive dead cells. FITC-labelled sheep anti-mouse antibodies and anti-human Fc antibodies (both Jackson Laboratories) were used for visualisation. (B) Expression of EphA3 in melanoma cell lines was examined by northern blot analysis. LK63 human B-cell leukaemia cells were used as positive control (lane1); LiBr cells (lane 2), AO2 cells (lane 3), AO9 cells (lane 4). The 28S mRNA is shown as loading control.