(A) HT29 cells were seeded on coverslips, labeled with antibodies against Dido3 and αTubulin, and visualized by confocal fluorescence microscopy. Mitotic phases represented are indicated on the left. Dido3 exclusion from the chromatin is first observed in prophase, is maintained while chromosomes are condensed, and finally reverts in telophase.
(B) HT29 cells were seeded on coverslips, labeled with antibodies against Dido3 and antibodies against H3K4me3, H3T3ph, or H3T6ph, and subsequently studied by confocal fluorescence microscopy. Dido3 loses affinity for condensed chromosomes, even though the H3K4me3 mark is maintained (left). Dido3is excluded from H3T3ph- and H3T6-positive chromatin (middle and right) in early prophase, before chromatin condensation is evident, and maintained in fully condensed chromosomes. Scale bars, 10 μm.
See also Figures S5 and S6.