Dictyostelium Carmil, but not Leep1, interacts with CPs. (A) Sequence alignment of Leep1 and Dictyostelium Carmil. Orange box, pink box, cyan box, and green underline indicate the verprolin-like region, acidic region, CP interaction motif, and proline-rich domain in Carmil, respectively. The red asterisk marks A736 in Leep1. (B) Sequence alignment of CP interactions from selected CARMILs. Cyan shading indicates residues that are identical to the consensus sequence shown at the top. (C) Immunoprecipitation performed using GFP-trap agarose. Samples were immunoblotted with anti-GFP or anti-CP antibody. (D) Time-lapse imaging of GFP-Carmil in vegetative cells. (E) Time-lapse imaging of GFP-Carmil in cells responding to cAMP stimulation in the absence or presence of LatA. Scale bar = 10 µm. C. elegans, Caenorhabditis elegans; IP, immunoprecipitation.