Figure 8. Filamin deficiency facilitates trafficking of lymphocytes.

(a; Left) Western blotting of lysates from f/f or −/− T cells with anti-FLNa or -b. Actin is a loading control. (Centre) The adhesive events of f/f and −/− T cells with the endothelium under shear flow. *¹P<0.001 versus total frequency of f/f T cells, *²P<0.001 versus total frequency of unstimulated −/− T cells,*³P<0.03 versus total frequency of stimulated −/− T cells. (Right) Effects of anti-LFA-1 on rolling velocities without CCL21. *¹P<0.01 versus f/f cells. *²P<0.01, compared with untreated −/− T cells. (b; Upper) Displacement (left) and velocity (right) of f/f or −/− T cells on ICAM-1 in the presence or absence of CCL21 (n=50). *¹P<0.02, *²P<0.05 versus f/f cells. (Lower) Each line represents a single-cell track of f/f or −/− T cells on ICAM-1 in the presence of CCL21. (c; Left) f/f or −/− T cells transmigrated through uncoated transwell in the presence of CCL21. (Right) f/f or −/− T cells transmigrated through ICAM-1-coated transwell is *P<0.002 versus corresponding f/f T cells. (d) f/f or −/− T cells labelled with CFSE or CMTMR were injected into normal mice. Flow cytometric profiles of blood and lymph nodes of the recipient after 1 h are shown. Numbers indicate the ratio of −/− to f/f cells. (Right) Ratios of −/− to f/f T cells are shown (n=4). *P<0.02 versus f/f T cells. (e; Left) Lysates from BAF/LFA-1/L-selectin cells introduced with Halo-tagged wild-type (WT) or Δ3 FLNa were immunoblotted with anti-Halo. (Centre) Adhesive events of those cells with the endothelium under shear flow. *¹P<0.03 versus total frequency of untreated WT cells, *²P<0.01, versus total frequency of the corresponding WT cells. (Right) KIM127 expression in those cells ±CXCL12 (n=5 of each group). *P< 0.05 versus WT FLNa-expressing cells. All data show the means±s.e.m. or representative of three independent experiments.