Extended Data Fig. 4: Bulk biomolecular condensate formation and quantification of condensate volume, condensed DNA length, and condensation probability.

a, Three per cent 30 K PEG triggers FoxA1 condensate formation in bulk at 50 μM in storage buffer: 20 mM HEPES (pH = 6.5), 100 mM KCl, 1 mM MgCl₂, 3 mM DTT, and 2 M Urea. b, Two per cent 30 K PEG triggers NH-FoxA1 condensate formation in bulk at 70 μM in storage buffer. c, The addition of 10 μM 32-BP ssDNA oligomers nucleated droplets of H1 in bulk at 90 μM that exhibited features of liquid-like droplets consistent with literature^28,29. These data demonstrate that H1-DNA form liquid-like condensates, which could be driven via transient crosslinking of H1 and DNA or H1-H1 interactions. Both mechanisms are accounted for in our free energy description. d, Condensate volume quantification of a representative time-averaged projection of a FoxA1-DNA condensate, where the black cross is the condensate peak location and the white dashed line is the intersecting profile to measure the volume. Lower panel: the black dots are the profile’s background-subtracted values and the solid black line is a Gaussian fit. The gray line represents the threshold value computed from the gradient of the Gaussian function that defines the edges of the condensate (see Methods). e, Condensed DNA length quantification of a representative time-averaged projection of FoxA1 and DNA. Below: the integrated one-dimensional DNA profile is defined into condensed versus non-condensed regions using the median of the profile’s median (gray) plus a tolerance (black dashed). f, Local correlation quantification of a representative time-averaged projection of FoxA1 and DNA. The condensates were localized (black crosses) and then 0.9 μm × 0.5 μm boxes centered around these peaks were cropped. The correlations between the cropped regions of FoxA1 (left) and DNA (right) were then computed.