Ephrin-B1 cytoplasmic tail tyrosines are not required for HSPG binding. A) Mock transfected (M) or HEK293T cells expressing ephrin-B1 (B1), ephrin-B1Q (B1Q), ephrin-B1S (B1S), or ephrin-B1QS, using PEI were 35S-sulfate labelled, lysed and precipitated with EphB2-fc. B) Flow cytometry of cells expressing ephrin-B1 (grey) or ephrin-B1S (black) incubated with EphB2-fc and PE-antibody. C) Flow cytometry of cells expressing ephrin-B1Q (grey) or ephrin-B1QS (black) after binding of EphB2-fc followed by PE. D) Western blot of lysates from mock (M) transfected and cells expressing human ephrin-B1 (B1), ephrin-B1S (B1S), ephrin-B1Q (B1Q), ephrin-B1QS (B1QS), or mouse ephrin-B1 (B1M). Upper panel detection with anti-ephrin-B1, lower panel with anti-actin for expression level control. Band regions were quantified and relative intensities compared to actin bands and related to lane 3 (insert).
