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. Author manuscript; available in PMC: 2010 May 1.
Published in final edited form as: Dev Cell. 2009 May;16(5):661–674. doi: 10.1016/j.devcel.2009.03.009

Figure 1. Polarized interaction between G-actin and Tβ4 in EC subjected to wound-induced migration.

Figure 1

(A) Preferential localization of G-actin and Tβ4 at the leading edge. G-actin and F-actin were visualized by immunofluorescence using Alexa Fluor 488-conjugated DNase I and Alexa 350-phalloidin, respectively. Tβ4 or Tβ10 were antibody-treated and visualized with Alexa 568-IgG. Direction of cell migration is shown. (B) Subcellular interaction between actinG13R and Tβ4 by acceptor photobleaching (AP) FRET. ECs were transfected with pGFP-actinG13R plus pDsRed-Tβ4 (left) or pGFP-DsRed (right, positive control). Cells were fixed and subjected to FRET assay. FRET efficiency (EFRET) in center and edge is shown (mean ± s.e.m., 8–12 cells). (C) Distribution of actinG13R–Tβ4 interaction in migrating ECs by sensitized emission FRET. ECs were transfected with pGFP-actinG13R plus pDsRed-Tβ4, pGFP plus pDsRed (negative control), or pGFP-DsRed (positive control), and EFRET analyzed with PFRET software (n = 6–10 cells).