Fig. 4.

p190 and anillin interact during cytokinesis. (A) p190-dependent multinucleation phenotype was rescued by the myosin II inhibitor blebbistatin. Failed cytokinesis events were measured by assessing the percentage of binucleated or multinucleated interphase cells relative to the total population of cells (n≥300). Results are mean±s.d. from three independent experiments. *P<0.005 for p190-siRNA-treated cells compare to Luc-siRNA-treated cells, and *P<0.03 for p190 siRNA compared to p190-siRNA-treated cells in 0.25 µM blebbistatin. (B) Co-immunoprecipitation of anillin and p190 in asynchronous or cytokinetic HeLa cells. HeLa cells were either untreated (Asyn) or synchronized with nocodazole (Cyto). Cells were then washed out of nocodazole and either mock-treated or treated with 50 µM blebbistatin for 40 minutes and cell lysates generated for immunoprecipitations (IPs). Western blots shown are representative of n>3. WCL, whole cell lysate. (C) Diagrams of p190 and anillin constructs used for recombinant protein expression. MBP and MBP-tagged C-terminal anillin (MBP-ANL) were bound to amylose beads, which were incubated with equal amount of 6×His tagged p190(1-763). Interaction was detected by western blotting with anti-His and anti-MBP antibodies.