FIGURE 1.

Arl4A is a novel ELMO-interacting partner. A, Arl4A^WT and Arl4A^Q79L interact with ELMO1 in a yeast two-hybrid system. Yeast strain EGY48 cotransformed with LexA BD fusion construct of ELMO1 and the B42 fusion constructs of the indicated Arl4s were grown on selective (−histidine, −tryptophan, −leucine) and nonselective (−histidine, −tryptophan) medium for a nutrient selective growth assay. B, Arl4A and ELMO1 interact in vivo in cells. HEK293T cells transfected with the indicated plasmids were subjected to a cross-linker, lysed, and immunoprecipitated (IP) with an anti-Myc antibody (ELMO1). Immunoblot analysis using anti-Myc and FLAG-HRP antibodies established the coprecipitation of ELMO and Arl4A proteins. C, Arl4A interacts with all forms of ELMO. HEK293T cells transfected with the indicated plasmids were cross-linked, lysed, and immunoprecipitated with an antibody against the Myc epitope (ELMO1–3). The coprecipitation of the various ELMO proteins and Arl4A was analyzed via immunoblotting with anti-Myc (ELMO1–3) and anti-FLAG-HRP (Arl4A) antibodies, respectively. TCL, Total Cell Lysate.