Rho activation promotes a monolayered cell division and polarity phenotype in WIF-B9 cells. (A–H) WIF-B9 cells untreated (control) and treated with a Rho activator were fixed and stained as indicated (A, C–E, and G). Architecture of 2D cultures (A and C), anaphase (D and G), and cytokinesis profiles (E) are shown. (A and C) Cells not contacting the substratum are denoted by an asterisk in x-z views. The percentage of cells contacting the substratum (B) was quantified. (A) Note the columnar-like pattern of ZO-1 distribution in Rho-activated WIF-B9 cells. (D) White and yellow arrowheads denote junctional-activated myosin II in the mitotic cell and its neighbors, respectively. (E–H) Comparison of the percentage of cytokinesis with both daughter cells contacting the substratum (E), anaphase β angle and m, h/d, Cx-z values (G and H, respectively), and EcadTSMod FRET efficiency (F). (A and C–E) Red arrowheads show hepatocytic lateral lumina. (E) Black arrowhead shows daughter that lost substrate contact. n = 20 cells/experiment were analyzed for N = 3 independent experiments (B and E–H). Error bars indicate ±SEM (dot graph) and +SD (bar graphs). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ns, nonsignificant, analyzed by t test.
