Extended Data Fig. 2: YAP does not form droplets in vitro and in vivo in the absence of crowding agents.

a, GFP-YAP purified from E. coil were analyzed by SDS-PAGE and visualized by Coomassie blue staining. b, GFP-YAP at varying concentrations was subjected to the droplet formation assay at room temperature and in the presence of 500 mM NaCl. c, 50 μM GFP-YAP was subjected to the droplet formation assay at room temperature in the presence of indicated salt concentrations. d, 50 μM GFP-YAP was subjected to droplet formation in the presence of 150 mM NaCl at 4 °C or 37 °C. e, Two YAP isoforms, GFP-YAP1–1β or GFP-YAP1–2α, did not form droplets (50 μM protein, 500 mM NaCl and room temperature). aa, amino acids. f, 50 μM GFP-YAP formed droplets in the presence of 10% PEG-8000, Ficoll or Dextran but not 10% glycerol or sucrose. Droplet formation assay was performed in the presence of 500 mM NaCl at room temperature. g, 50 μM GFP-YAP did not form droplets in the presence of BSA at varying concentrations. h, GFP-YAP did not form nuclear puncta in both HeLa cells and 293T cells. Scale bars, 10 μm. Experiments in a–h were repeated independently three times with similar results.