Fig. 5: Phase separation of Taf14_ET.

a Representative fluorescence microscopy images of the Taf14_ET–GFP fusion protein at different concentrations. The droplet formation buffer comprises 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% PEG8000. The scale bars indicate 10 µm; note that the same scale applies to all of the panels. b Representative images of Taf14_ET-GFP at different time points showing the dynamic fusion of two droplets in the droplet formation buffer. c Representative images from FRAP experiments confirm the fluidity of Taf14_ET-droplets. d Quantification of the average intensity of recovered fluorescence in the bleached region of droplets at 10 s intervals. All of the data are presented as means ± SDs (n = 16). Source data are provided as a Source Data file. e Turbidity measurements as determined by light scattering at 350 nm. Sumo-Sth1_EBM significantly increases the turbidity of GST-Taf14_ET at 10 and 25 μM protein concentrations. Error bars represent standard deviations of three replicates. NS for P > 0.05; *** for P < 0.001; **** for P < 0.0001; two-tailed Student’s t-test. Data are presented as mean ± SD, n = 3. f Representative images of the mixture of Taf14_ET-GFP (5 µM) and Sth1_EBM-RFP (5 µM) in the droplet formation buffer showing Sth1_EBM increased the phase separation of Taf14_ET. As controls, RFP or Sth1_EBM^M4-RFP did not affect the phase separation of Taf14_ET. g Representative images of the mixture of Taf14_ET-GFP and Sth1_EBM-RFP or Snf5_EBM-CFP in the droplet formation buffer. Both Sth1_EBM and Snf5_EBM strongly promoted the phase separation of Taf14_ET compared with the addition of RFP. h Representative images of a mixture containing the Taf14_ET-GFP, Sth1_EBM-RFP, and Snf5_EBM-CFP fusion proteins in droplet formation buffer. Note that all droplets contain all three fluorescence signals, indicating colocalization of three proteins.