Western blot analysis with ephrin-A4 (s) specific antiserum.
(A) COS cells were transfected with ephrin-A4 (m) cDNA or ephrin-A4 (s) cDNA, and cells and culture supernatant were harvested 3 days posttransfection. Cells were lysed. Either 10 μg protein was applied in each lane (left panel), or 10 μL culture supernatant was applied in each lane (right panel). (B) Tonsil T cells or tonsil B cells were isolated and lysed, and 10 μg of protein was applied in each lane. (C) 105 blood B cells were stimulated for 6 days, as indicated in the figure text. Control is no stimulation. The culture supernatant was harvested, and 10 μL was applied in each lane. All blots were stained with ephrin-A4 (s) specific polyclonal antiserum. The arrowhead denotes the ephrin-A4 (s) protein band. Molecular weight in kDa is indicated to the left of each panel. Tsup: supernatant of PHA-stimulated pooled T cells.
