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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.