Fig. 9. Subcellular localization of MLCK, ROCK and myosin compared to actin in T cells migrating on ICAM-1. MLCK (A) and ROCK (B) were detected with mAbs specific for MLCK and ROCK I, followed by Alexa 488-goat anti-mouse IgG and counter-stained with Alexa 546-phalloidin to visualize F-actin. Images are displayed as gradient maps where red represents high and blue low fluorescence intensity. (C) Myosin (green) and F-actin (red) were merged to determine colocalization (yellow) within an Mg²⁺/EGTA-stimulated T cell on ICAM-1. (A,B) Images represent projections onto the x-y plane of all individual optical sections taken along the z-axis using maximal fluorescence values. Image C represents a single section in the x-y plane.