Fig. 3
From: Unraveling the molecular interactions involved in phase separation of glucocorticoid receptor
Relation between the formation of GR condensates and chromatin. a U2OS cells expressing GFP-GR were incubated with Dex and time-lapse imaged by confocal microscopy. 2D trajectories of GR foci were obtained by single-particle tracking. Red arrows point to foci very close to nucleoli, blue arrows show foci far from these structures (Scale bar: 5 μm). (Left panel) Representative trajectories obtained from these analyses. (Middle panel) The mean square displacement (MSD) was calculated as a function of the time lag (τ) according to Eq. 3. MSD(τ) data obtained for representative trajectories of foci close to nucleoli (red, proximal) or far from them (blue, distal). (Right panel) The mean explored distance (MEDτ = 100) was calculated from the MSD values at τ = 100 s. An average MEDτ = 100 was calculated for the proximal and distal populations of foci. The bar marked with an asterisk (*) represents data significantly different (p < 0.05) (Proximal: nfoci = 30; Distal: nfoci = 50; ncells = 7). Raw data can be found in Additional file 15: Supplementary Table S3. See also Additional file 7: Supplementary Video S3. b, c U2OS cells co-expressing GFP-GR and H2B-mCherry were incubated with Dex and imaged by confocal microscopy (Scale bar: 5 μm). b The zoomed image corresponds to the region indicated by the dashed square (Scale bar: 2 μm). c U2OS cells were incubated 16 h with TSA 1 μg/ml before Dex stimulation (Top panel) GR foci density relative to the mean density of cells only treated with Dex (Dex: ncells = 82; Dex + TSA: ncells = 71). (Bottom panel) Coefficient of variation (CV) of H2B-mCherry intensity in the nucleus (Dex: ncells = 25; Dex + TSA: ncells = 18). The bar marked with an asterisk (*) represents data significantly different (p < 0.05) from cells only treated with Dex. Raw data can be found in Additional file 15: Supplementary Table S3