Canoe regulates Pins-mediated spindle orientation. (A–G) S2 cell spindle orientation assay. Representative images are shown, and the quantification of each experiment is shown in H. Ed:GFP or Ed:GFP:Pins proteins were induced to form cortical crescents by cell aggregation, and the angle of the mitotic spindle was measured relative to the center of the cortical crescent. Pins cortical localization (green), mitotic spindle (α-tubulin [αTub]), and merge (in some cases also showing the mitotic DNA marker phospho–histone H3 [PH3]) are shown. CDS, coding sequence. Bar, 5 µm. (H) Quantification of experiments shown in A–G depicted as a cumulative plot. Random spindle orientation is a diagonal line (e.g., Ed:GFP); optimal spindle orientation is reflected in a leftward shift in the plot (e.g., Ed:Pins^TPR+LINKER), and partial spindle orientation falls in between. The key is an abbreviated version of the experiments shown on the left in A–G. CnoCDS RNAi, n = 36; Cno 3′ UTR RNAi, n = 36; CnoFL Rescue, n = 30; CnoDelta RA Rescue, n = 30; Cno RNAi + Mud RNAi, n = 29; Ed:PinsTPR+linker, n = 30; Ed:GFP, n = 33.