Kank regulates RhoA-dependent formation of actin stress fibers and cell migration via 14-3-3 in PI3K-Akt signaling -- Data Supplement - JCB--Kakinuma et al. -- The Journal of Cell Biology

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Figure S4. The interaction between Kank and IRSp53 is only slightly affected by 14-3-3. (A) Kank as well as Kank^S167A bind to IRSp53. HEK293T cells expressing GFP-IRSp53 (lanes 3 and 4) or control GFP (lanes 1 and 2) along with Kank-FLAG (lanes 2 and 3) or Kank^S167A-FLAG (lane 4) were subjected to immunoprecipitation using an anti-FLAG antibody. (B) 14–3-3θ does not interfere with the interaction between MBP-coil and GST-IRSp53. MBP-coil was subjected to a pull-down assay in vitro using GST-IRSp53 in the presence of various amounts of His-tagged 14-3-3θ (lanes 3–5). MBP-coil^S167A was also examined (lane 6). MBP, which was pulled down with GST (lane 1) or GST-IRSp53 (lane 2), was used as a control..