Slower migration of ezrin T567E transgenic T cells in lymph node in vivo. One day before imaging, WT B-lymphocytes, which were stained with blue (CMF2HC), were injected to delineate B-cell follicles. T-lymphocytes from WT mice were stained red (CMTMR) and from ezrin T567E transgenics were stained green (CMFDA), mixed, injected intravenously, and migration in inguinal lymph node was assessed 18 hours later by 2-photon microscopy. (A) Image of region of central T-cell cortex marking representative region for analysis. Bar represents 100 μm. (B) Frame from region analyzed along with tracks of WT (green) and ezrin T567E transgenic (red). Bar represents 30 μm. (C) Summary of migration speed of individual cells shows lower average speed of ezrin T567E transgenic lymphocytes (P < .0001). Data are mean ± SEM: WT (10.8 ± 0.3 μm/s; N = 55), Tg (8.7 ± 0.4 μm/s; N = 40). (D-F) Comparable analyses of migration near the border between T-cell area and B-cell follicle. Migration speed mean ± SEM: WT (12.2 ± 0.5 μm/min; N = 39), Tg (6.4 ± 0.3 μm/min; N = 30). All P values were calculated using the Mann-Whitney test in Prism Version 5 (GraphPad). ***P < .0001.
