Extended Data Fig. 2: Time-lapses of cell cycle regulated proteins.

a, The specificity of the anti-ANLN antibody was shown by co-localization with mNG-tagged ANLN. Scale bars, 10 μm. b, Time-lapse microscopy for mNG-tagged ANLN protein demonstrates an increase in nuclear expression over the course of the cell cycle with a peak in late G2. During mitosis and cytokinesis, ANLN localizes to the cell membrane and is involved in the formation of the cleavage furrow (t = 17–18 h and t = 35 h). Scale bar, 10 μm. c, The specificity of the anti-AURKB antibody was validated by co-localization with mNG-tagged AURKB. Scale bars, 10 μm. d, Time-lapse microscopy for mNG-tagged AURKB protein demonstrates an increase in protein expression over the course of the cell cycle with a peak in late G2. During mitosis, AURKB localizes to the mitotic chromosomes at hours 13 and 29. Scale bars, 10 μm. e, The specificity of the anti-GATAD1 antibody was validated by co-localization with mNG-tagged GATAD1. Scale bars, 10 μm. f, Time-lapse microscopy for mNG-tagged GATAD1 protein demonstrates a decrease in nuclear abundance throughout the cell cycle (mitosis at 35 h). Scale bars, 10 μm. g, Time-lapse microscopy for mNG-tagged KIFC1 protein demonstrates a CCD translocation from the nucleoplasm to the kinetochores in late G2 into mitosis at hours 14 and 30 and in early G1. Scale bars, 10 μm. h, Time-lapse microscopy for mNG-tagged NET1 protein demonstrates a CCD increase in protein abundance and translocation from the nucleoplasm to the nucleolus (mitosis at 35 h). Scale bars, 10 μm.