Maximum intensity projections of guard cells from different angles with centromere labeled by HTR12-Venus in green and chromatin labeled by Histone HTB1-CFP in magenta (see also Supplemental Video 1). (A) Projection image of guard cells from 20 image sections with z-interval 0.2 μm. Centromeres in one guard cell were arbitrarily numbered from 1 to 10. (A′) Diagram of positions of centromeres in A. The *x*-axis represents cell wall between two guard cells, the *y*-axis is vertical to the *x*-axis, and the *z*-axis is directed to the bottom of the guard cell nuclei. Centromeres were defined as brightest spot in the local fluorescent locus. Centromeres from sections 1–10 are represented by dark gray-scaled dots and highlighted by numbered arrows (1, 2, 4, 5, and 10). Centromeres from sections 11–20 are represented by light gray-scaled dots and highlighted by numbered arrows (3, 6, 7, 8, and 9). Also see Supplemental FigureS1. (B) Projection image of guard cells after 30° counterclockwise rotation along vertical axis. Centromeres (3, 6, 7, and 9) at the bottom side of the nuclei move left. Centromere 9 can be observed at the periphery of the projected nuclei. Centromeres (4 and 10) at the topside of the nuclei move right. (C) Projection image of guard cells after 60° counterclockwise rotation along the vertical axis. (D) Projection image of guard cells after 90° counterclockwise rotation along the vertical axis. Centromere 4 and centromere 7, which were at the center of the projected image at 0° (A), can be observed at the nuclear periphery of the projected nucleus. (E) Projection image of guard cells after 120° counterclockwise rotation along the vertical axis. (F) Projection image of guard cells after 150° counterclockwise rotation along the vertical axis. (G) Projection image of guard cells after 180° counterclockwise rotation along vertical axis. Bar, 5 μm.

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