Fig. 3

Recruitment of Syk kinase to FcγR-IgG microclusters. a Syk (green) associated with FcγR-IgG clusters (magenta) at the leading edge and during retrograde movement of the FcγR-IgG; scale bar 5 μm. B-C) Magnified region from (a) shows the association of Syk with FcγR-IgG clusters at the pseudopod leading edge (b) and retrograde movement following their detachment from the leading edge (c); scale bar 4 μm. d Tracking of individual FcγR-IgG microclusters from the cell in (a) (green indicates movement away from the cell center and red indicates movement toward the center of the cell); scale bar 10 μm. Intensity of IgG (e) and Syk kinase (f) associated with a single FcγR-IgG cluster for one track; color-coding indicates directionality of the FcγR-IgG microcluster. The black line indicates the local background signal. These results illustrate the typical behavior of FcγR-IgG microclusters as the they form at the leading edge followed by a static intensity; whereas fluctuations in Syk localization and an overall decrease in signal was observed as the microcluster departed from the leading edge. We performed tracking on one representative cell with a total number of 117 tracks. g Montage showing the oscillations in Syk intensity on the FcγR-IgG cluster; the width of the montage panel 2 μm