Fig. 2
From: Unraveling the molecular interactions involved in phase separation of glucocorticoid receptor
GR condensates are modulated by osmolarity changes. a, b U2OS cells expressing GFP-GR were incubated with Dex, then with medium supplemented with 100 mM NaCl for 1 min and imaged by confocal microscopy (Scale bar: 5 μm). a Representative images of the same cell before and after NaCl incubation, and after re-introducing the cells in isotonic medium (NaCl washout). (Top panel) Mean foci density relative to the density obtained before NaCl incubation in the same cells. (Bottom panel) Mean foci intensity relative to the intensity of the nucleus in each image. Bars with different superscript letters indicate data significantly different (p < 0.05) (ncells = 5). Raw data can be found in Additional file 14: Supplementary Table S2. b Representative images of the same cell before and after incubation with NaCl at the indicated time points. Mean foci density (red line) and mean foci intensity (blue line) (ncells = 4). Foci intensity relative to the intensity of the nucleus in each image. Foci density relative to the density before NaCl incubation. Raw data can be found in Additional file 14: Supplementary Table S2. c U2OS cells co-expressing GFP-GR and H2B-mCherry were incubated with Dex, then with medium supplemented with 100 mM NaCl for 5 min and imaged by confocal microscopy. Representative images of the same cells before and after with NaCl (Scale bar: 5 μm). Intensity profiles obtained along the lines indicated with gray lines in the merged images. The intensity is represented relative to that determined in the nucleoplasm. Blue arrows point to the position of GR hyperosmotic-induced dots (HOIDs), which are excluded from chromatin condensates. This experiment was run in 12 cells obtaining a similar anticolocalization pattern. d U2OS cells co-expressing H2B-mCherry and GFP-GR or GFP were incubated with vehicle (Veh), then with medium supplemented with 100 mM NaCl for 1 min and imaged by confocal microscopy. Representative images of cells before and after NaCl incubation (Scale bar: 5 μm). This experiment was run in six cells for each condition obtaining similar distributions