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Figure 6


Fig. 6. Signaling through JAM-A, PI 3-kinase and PKC is required for JAM-A-induced migration. (A) Western blot of protein lysate (50 µg) from mock-, JAM-A- and JAM-A {Delta}-257-transfected HUVECs, indicating level of expression of JAM-A. [Note: deletion of the cytoplasmic domain ({Delta}-257) results in a lower-sized band.] (B) Cell migration on vitronectin using cells as in A. (C) Western blot showing the absence of HA-tagged JAM-A {Delta}-257 in the immunoprecipitate (IP) of anti-{alpha}vß3 from total cell lysate (Input) of serum-starved HUVECs transfected with the JAM-A {Delta}-257 construct and treated with or without RGDS as indicated. Isotype-specific IgG was used as a negative control (cIgG), and the blot was reprobed with anti-ß3 antibody, to ensure equal loading. Mock-transfected or JAM-A-overexpressing cells were pre-treated with various concentrations of anti-JAM-A antibody (D), or with DMSO or wortmannin or Bis (E) for 20 minutes and then assayed for migration on vitronectin. *P<0.05.





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