Figure 10. The F0, F1 and F3 domains of Kindlin‐2 are essential for connecting the cadherin/catenin complexes to the actin cytoskeleton and AJ stabilization.

A, WT and various deletion mutants of Kindlin‐2 tagged to EGFP, as well as control EGFP, were expressed in HEK293 cells and immunopurified from resting confluent cell monolayers with EGFP‐Trap Sepharose. The immunoprecipitates were analysed on western blots with the indicated Abs. B, β‐catenin‐GST was incubated with lysates of the HEK293 cells expressing WT and Kindlin‐2 deletion mutants. Kindlin‐2 mutants were purified with EGFP‐Trap Sepharose and analysed on western blots with Abs to GST and EGFP. The values in the western blots indicate band intensities expressed as a percentage of control cells expressing full‐length Kindlin‐2, which were assigned a value of 100%. C, HUVECs were treated with non‐targeting or Kindlin‐2‐siRNA (100 nm) and, 24 h later, were transfected with full‐length Kindlin‐2‐EGFP or its mutants, as indicated. TEER changes were monitored in the absence or presence of thrombin (4 U ml⁻¹) 24 h post‐transfection. Data are the mean ± SEM. ^* P < 0.01 (n = 6), ΔF0, ΔF1 or ΔF3‐Kindlin‐2‐transfected HUVEC vs. EC transfected with full‐length Kindlin‐2.