Figure 5.

Focal adhesion proteins are recruited to sites of integrin engagement. (A) Primary human T cells were stimulated on coverslips patterned with ICAM-1 surrounded with OKT3 for 15 min, and labeled with m24 to detect the active, extended-open conformation of LFA-1, and Kim127 to detect the extended form of LFA-1. (B,C) Jurkat T cells were stimulated on VCAM-1 patterns surrounded with anti-CD3 for 15 min. Cells were then fixed and labeled with phalloidin to detect F-actin, and with antibodies to talin, vinculin, and paxillin. (D,E) Primary human T cells were allowed to interact with VCAM-1 (D) or ICAM-1 (E) patterns surrounded with OKT3 for 17 min. Cells were then fixed and labeled with phalloidin to detect F-actin, and with antibodies to talin and vinculin. Far right panels in (B–E) show cropped regions in which the four channels have been merged. (E) Scale bars = 10 µm.