Tilted metaphase spindle alignment occurs during anaphase in columnar- but not in hepatocytic-polarized Par1b cells. (A–C) MDCK-Par1b cells uninduced (+Dox) or overexpressing Par1b (−Dox) were fixed and stained as indicated. Asterisks in A and black arrowhead in B indicate nonsubstrate contacting cells and daughters that lost substrate contact, respectively. The percentage of cytokinesis with both daughter cells contacting the substratum (B) and the metaphase and anaphase β angle (C) were quantified. n = 20–25 cells/experiment were analyzed for N = 3 independent experiments. Error bars indicate +SD (bar graphs) or ±SEM (dot graph). *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ^§§, P ≤ 0.01; ^§§§, P ≤ 0.001; ns, not significant, analyzed by t test. (D) x-z-t Confocal time-lapse sequence of dividing MDCK cells overexpressing Par1b with columnar phenotype. The β angle was followed in seven cells from metaphase to anaphase/telophase profiles. (B and D) Dotted line show cell contours. (A–D) Red arrowheads show hepatocytic-like lateral lumina.