FIGURE 5.

Immunostaining and FRI of focal adhesion proteins. REF52 cells transfected with YFP-paxillin were fixed and immunostained with primary antibody against vinculin, followed by Cy5-conjugated secondary antibodies. Actin filaments were visualized with phalloidin-TRITC. Cells on 58-nm and 108-nm RGD-nanopatterns were observed at 3 h and 24 h after plating. The rows present the images with paxillin in red, vinculin in blue, and actin in green. The last row shows the ratio between paxillin and vinculin intensities. FRI are presented in a spectrum scale as indicated in the lookup table.