Figure 2.

Plasma Membrane Association of ECT2 in Anaphase Is Required and Sufficient to Support Cytokinesis

(A) Synchronization scheme for anaphase-specific membrane targeting of ECT2-C1B proteins.

(B) Live-cell imaging of ECT2-C1B and ECT2-C1B^Q27G cell lines. Cells were transfected with ECT2 siRNA and synchronized in metaphase using the protocol depicted in (A). Cells were treated with DMSO or TPA 45 min after release from metaphase and imaged using bright-field microscopy. Time point t = 0 min was set to the metaphase-to-anaphase transition.

(C) Quantification of cytokinetic phenotypes of cells recorded in (B). Mono-nucleated cells that were in metaphase at the beginning of recording were scored (n > 200 each, bars represent mean ± SD of three independent experiments, Student’s t test).

(D) Synchronization scheme for membrane targeting of ECT2-C1B proteins in metaphase followed by TPA washout before anaphase onset.

(E) Quantification of cytokinetic phenotypes of cells treated as shown in (D). Mono-nucleated cells that were in metaphase at the beginning of recording were scored (n > 65 each, bars represent mean ± SD of three independent experiments).

See also Figure S2.