Ile 400

Cells were placed in a 37 °C humid incubator by controlling the temperature and CO₂ flow using H201-couple with temperature and CO₂ units. Live chromatin imaging was performed using a DMI8 inverted automated microscope (Leica Microsystems) featuring a confocal spinning disk unit (CSU-X1-M1N, Yokogawa). An integrated laser engine (ILE 400, Andor) was used for excitation with a selected wavelength of 647 nm and 140 mW as excitation power. A 100× oil immersion objective (Leica HCX-PL-APO) with a 1.4 NA was chosen for a high-resolution imaging. Fluorescence emission of the SiR-Hoechst was filtered by a single-band bandpass filter (FF01-650/13-25, Semrock, Inc.). Image series of 150 frames (5 fps), with exposure time of 150 ms per frame, were acquired using Metamorph software (Molecular Devices) and detected using sCMOS cameras (ORCA-Flash4.0 V2) and 1 × 1 binning, with sample pixel size of 65 nm. All series were recorded at 37 °C.

Vegetative DicMaLionR cells were seeded on a 35‐mm glass‐bottom dish (P35G‐1.5‐14‐C, MatTek, Ashland, MA, USA) and washed twice with KK2 buffer after attachment, then filled with KK2 buffer for observation. Five‐minutes later from the start of observation, oligomycin (mixture of A, B, and C isomers; O‐500, Alomone Labs, Jerusalem, Israel), an inhibitor of ATP production, was added at the final concentration of 2.5 μM. Time‐lapse images were acquired every 10 s using 60× oil immersion (PlanApo 60×/1.40 NA, Nikon, Tokyo, Japan) objective lens and Dragonfly200 (Andor, Tokyo, Japan) equipped with EMCCD (iXonUltra 888, Andor, Tokyo, Japan) camera. A solid‐state laser (ILE‐400, Andor, Tokyo, Japan) was used as the source for providing 561‐nm (RFP excitation) wavelength lights.