Cdr2 N-terminal clustering is under the control of Pom1 and Cdr2 kinase activity. (A) Medial plane epifluorescence images of Cdr2-mEGFP and Cdr2N-Kcc4C–GFP chimera in wild-type (wt) cells or cells expressing Pom1-chimera or carrying a Cdr2 kinase-dead mutation (KD; E177A). (B) Coimmunoprecipitation assays between Cdr2N-Kcc4C–GFP and Mid1 in wild-type cells and cells expressing the Pom1-chimera or Cdr2N^KD-Kcc4–GFP and Mid1. Immunoprecipitations were performed with anti-GFP mAb. Inputs and immunoprecipitation (IP) samples were probed with anti-GFP mAb and anti-Mid1 affinity-purified antibody. Mean normalized Mid1 coimmunoprecipitation signals from four independent experiments are shown on the right. Error bars show SDs. Molecular masses are indicated. (C, top) Medial plane confocal images of Mid1-mEGFP and Cdr2-TagRFP in wild-type cells (left) and cells expressing the Pom1-chimera (center) or of Mid1-mEGFP and Cdr2^KD-TagRFP (right). (bottom) Mid1-mEGFP and Cdr2-TagRFP intensity along the medial cortex in the same cell. (bottom right) Mean percentage of Cdr2-TagRFP nodes containing Mid1-mEGFP in individual cells. Error bars show SDs (n ≥ 25 cells). WB, Western blot. Bars, 5 µm.