Fig. 6: JMJD3 IDR is essential for condensate formation.

a mEGFP–JMJD3 and mEGFP–JMJD3 ΔIDR expression vectors were transfected into HEK293T (0.05ug). 24 h later total protein extracts were prepared and the JMJD3 (HA) and TUBULIN levels were determined by immunoblot. The image shown is representative of two independent experiments. Source data are provided as a Source Data file. b mEGFP, mEGFP–JMJD3 and mEGFP-JMJD3 ΔIDR proteins were analyzed using droplet-formation assays in nuclear extracts at room temperature in the presence of 150 mM NaCl. Quantifications of the droplets are displayed on the right. Data are the mean ± SEM. Boxes comprise values from Q1 to Q3 of the dataset; line corresponds to median value; whiskers show the data range (from min. to max. values within dataset). ***p < 0.001 (P values were calculated using one-tailed Student’s t test, p = 0.000113231). Droplets in 5 fields in each group from three biologically independent experiments were quantified. Scale bar, 5 μm. Source data are provided as a Source Data file. c Confocal microscopy images of HEK293T cells transfected with 0.05 ug mEGFP-JMJD3 or mEGFP-JMJD3 ΔIDR. The images are representative of three biologically independent experiments. Quantifications of the number of JMJD3 puncta are shown on the right. Data show the mean ± SEM. Boxes comprise values from Q1 to Q3 of the dataset; line corresponds to median value; whiskers show the data range (from min. to max. values within dataset). ***p < 0.001 (P values were calculated using one-tailed Student’s t test, p = 2.5178E-09). n = 20 transfected cells in each group were quantified. Scale bar, 5 μm. Source data are provided as a Source Data file.