Fig. 6
From: Unraveling the molecular interactions involved in phase separation of glucocorticoid receptor
Structural determinants involved in GR condensates formation. a, b Representative images of U2OS cells expressing GFP fused to wild-type GR or mutant variants of the receptor (GR407C, GRN525, GRtetra, GRmon) incubated with Dex, before (top panels) and after incubation with medium supplemented with 100 mM NaCl (bottom panels) (Scale bar: 5 μm). The ability of the different mutants to form foci in isotonic (light green) and hypertonic (dark green) conditions was evaluated by calculating the foci density (ncells,GR + Dex = 146; ncells,GR + Dex + NaCl = 15; ncells,407C + Dex = 43; ncells,407C + Dex + NaCl = 24; ncells,N525 + Dex = 27; ncells,N525 + Dex + NaCl = 16; ncells,tetra + Dex = 21; ncells,tetra + Dex + NaCl = 12; ncells,mon + Dex = 24; ncells,mon + Dex + NaCl = 18). Bars with different superscript letters represent data significantly different (p < 0.05). The asterisk (*) denotes a significantly different foci density (p < 0.05) with respect to that obtained for the same GR variant in isotonic medium. Raw data can be found in Additional file 17: Supplementary Table S5. c Representative images of U2OS cells co-expressing GFP-GR and mCherry-GRmon incubated with Dex and then with medium supplemented with 100 mM NaCl when indicated (Scale bar: 5 μm). Intensity profiles obtained along the gray lines indicated in the merged images. The intensity is represented relative to that determined in the nucleoplasm in each case. Similar results were obtained with U2OS cells coexpressing GFP-GRmon and mCherry-GR (ncells,Dex = 10; ncells,Dex + NaCl = 13, including both GFP-GR + mCherry-GRmon and GFP-GRmon + mCherry-GR experiments)