Fig. 5. Liquid-liquid phase separation of BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL hetero-dimer.

A Co-localization analysis of BCOR-mCherry with KDM2B-EGFP in live HeLa cells. B Co-localization analysis of BCOR-mCherry, PCGF1-ECFP and KDM2B-EGFP in live HeLa cells. C Observation of condensates for BCOR^{ANK-linker-PUFD} (wild type or mutant)/PCGF1^RAWUL in HeLa cells. WFY indicates the 5 aromatic residues on linker of BCOR, including W1598, F1600, Y1601, Y1614 and Y1633. D Effect of CaCl₂ and NaCl on the phase separation of BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL hetero-dimer with concentration of 60 μM. E Representative fusion event of BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL droplets. F In vitro FRAP analysis of the droplet of BCOR^{ANK-linker-PUFD}/PCGF1^RAWUL. G Normalized FRAP recovery curves for BCOR^{ANK-linker-PUFD-EGFP}/PCGF1^RAWUL droplet in (F). Data are mean ± SEM at each time point and combined data from three independent repeats. H Plasmids expressing BCOR^{ANK-Linker-PUFD} or PCGF1^RAWUL were co-transfected into HeLa cell. Before imaging for LLPS, cells were pretreated with 10 μM ionomycin and 5 mM CaCl₂ for 2 h. To observe the effect of calcium-chelator BAPTA-AM on the ionomycin induced LLPS, culture containing 10 μM ionomycin and 5 mM CaCl₂ was discarded, the cell was washed with PBS, then culture containing 10 μM BAPTA-AM was added to the cell.