JCS008607 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Phase-contrast micrograph of dissected mesentery from uninjected mice (left panel). Appearance of cells cultured for 20 hours after collection from mesenteric sheet is shown in the middle panel. Mesentery-derived cells were subjected to immunoblotting with anti-EphB2 or anti-EphB4 antibody (right panel). These cells express ephrin receptors EphB2 32 and EphB4.

• Supplemental Figure S2 -

  Fig. S2. The 35 kDa ectodomain fragment of ephrin-B1 inhibited the motility, but not the proliferation of EphB2-expressing cells. (A) COS-1 cells were transiently transfected with ephrin-B1 or control mock plasmid. Parent L cells or L cells stably expressing EphB2 were plated into Transwell inserts (8 μm pore) in conditioned medium of transfected COS-1 cells or medium containing 2 μg/ml of ephrin-B1-Fc or control Fc. The undersides of the insert membranes were coated with fibronectin. After incubation for 4 hours, cells that migrated through the membrane to the bottom surface were stained with Giemsa’s solution and counted. (B) Parent L cells or L cells stably expressing EphB2 were incubated with ephrin-B1-Fc or control Fc (2 μg/ml), or the conditioned medium of ephrin-B1 or mock-transfected COS-1 cells for 20 hours. Proliferation of the cells was evaluated by measurement of DNA synthesis through detection of BrdU incorporation by ELISA assay (Roche). The results from three independent experiments, each in duplicate, are shown as the mean ± s.d. *P<0.1.