Impaired integrin adhesiveness of Kindlin-3–deficient effector T cells in vitro. (A) Deficient Kindlin-3 expression on T effectors derived from CD4CrexKindlin-3^fl/fl mice and their control Kindlin-3^fl/fl littermates. Lysates were immunoblotted with anti–Kindlin-3 Ab or anti-tubulin Ab. (B) FACS staining of the major integrin subunits critical for lymphocyte-endothelial interactions and of the inflammatory G-protein coupled receptor CXCR3 on wt and Kindlin-3–deficient effector T cells. (C) Resistance to detachment of Kindlin-3–deficient or wt effector T cells by the indicated shear forces after settling for 1 minute under shear-free conditions on ICAM-1 or on VCAM-1 coated at 760 sites/μm². Results are the mean ± range of 2 measurements. *P < .05. (D) Resistance of wt and Kindlin-3–deficient T cells accumulating on low-density P-selectin (left panel) or E-selectin (right panel) to detachment by progressively elevated shear stresses. T cells were allowed to accumulate for 40 seconds on low-density P- or E-selectins (coated at 95 and 25 sites/μm², respectively) at a shear stress of 1 dyn/cm², and the flow was increased by stepwise increments every 5 seconds. The number of cells bound at the end of each interval was determined in 2 fields and was expressed as a percentage of initially accumulated lymphocytes. A representative of 3 experiments is shown. (E) Chemotaxis of Kindlin-3–deficient and wt effector T cells determined in a transwell assay. Unless indicated, all chemokines were placed at 10 nM at the lower well. Pore size: 5 µm. Results are the mean ± standard error of the mean (SEM) of 4 to 6 independent measurements.