Liquid-liquid phase separation (LLPS) drives chromocenter clustering in the DNA hypomethylated cells.
(A): Series of time-lapse 3D reconstruction images of the Dnmt1-KO ESCs expressing hKO1-HP1γ. Yellow squares indicate the region highlighted at the bottom panel. White arrows indicate the chromocenters undergoing the fusion. Images were captured and cropped from S1 Movie. Scale bar, 3 μm. (B): Representative immunostaining images of surface-spread nuclei of the Dnmt1-KO ESCs with or without 1,6-hexanediol (HD) treatment. About 10% of 2,5- or 1,6-HD were treated for 1 min. For the recovery experiment, cells were incubated in an ESC culture medium for 1 h. Scale bar, 5 μm. (C): Boxplot showing the number of distinct chromocenters in the nuclei of the Dnmt1-ESCs in the individual condition, related to (B). No treatment, n = 71; 2,5-HD treatment, n = 222; 1,6-HD treatment, n = 217; 1,6-HD treatment with recovery culture, n = 199. P values were calculated using the Mann-Whitney U-test. ***P < 0.001.