Classical cadherin-mediated adherens junctions regulate nucleus and centrosome positioning and cell orientation. (A–C) Astrocytes or JEG3 epithelial cells were plated onto fibronectin-printed micropatterns in the presence of 1.8 mM calcium. Cells were incubated for 2 h in presence of calcium and then for 5 h in calcium-free medium (−Ca), or incubated for 2 h in the absence of calcium and then for 5 h in calcium-containing medium (+Ca). (D–I) 3 d after nucleofection of the indicated siRNAs and pEGFP constructs, astrocytes and U373 astrocytoma cells were either plated onto fibronectin-printed micropatterns (immobile cells) or submitted to a wound-healing assay (migrating cells). (A, D, and G, top) Pan-cadherin or E-cadherin (green), pericentrin (red, white arrowhead), and Hoechst (blue) stainings. (A, bottom) Higher magnification of cadherin staining at cell–cell contacts. Bars, 20 µm. (B, E, and H) Distances between the nucleus and cell centroids (black) and between the centrosome and cell centroid (white). Data are given as mean ± SD of three independent experiments totaling at least 120 cells. (C, F, and I) Distribution of the angle between the centrosome–nucleus axis and the micropattern radius passing through the centrosome. The median angle is indicated in red. Statistical differences are indicated. *, P < 0.05; **, P < 0.005.