Fig. 5.

Rab11 traffics to the furrow on flagella and is required for abscission. (A) mNG-Rab11 localization through the cell cycle. In late telophase, Rab11 is recruited to intracytoplasmic axonemes (yellow arrowhead), and, immediately before anterior-posterior furrow ingression, Rab11 delineates the furrow (white arrow). mNG-Rab11 remained in the cleavage furrow and at the leading edge of the advancing furrow during cytokinesis (white arrow; time stamp, min:s). (B) Immunofluorescence localization of tubulin (grayscale), actin (green), HA-Rab11 (red), and DNA (blue). HA-Rab11 localized to the ER in interphase cells. During late telophase, Rab11 loaded onto flagella (yellow arrowheads). Just ahead of anterior-to-posterior furrow progression, HA-Rab11 accumulated in the furrow. Enlarged images correspond to cyan boxes with two or one optical section that show Rab11 and actin accumulating at the ends of the nascent flagella (blue arrowheads). (C) Rab11 antisense morpholino-treated cells are delayed in abscission and frequently have abnormal furrow positioning. (D) Histogram of division timing for Rab11 KD (n = 80) and morpholino control (n = 141). (E) Example of an actin-KD cell with abnormal posterior morphology and a membrane accumulation. (F) Quantification of this phenotype for actin depleted (n = 602) and morpholino control cells (n = 613). (Scale bars: 5 μm.)