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Fig. 1 | BMC Biology

Fig. 1

From: Unraveling the molecular interactions involved in phase separation of glucocorticoid receptor

Fig. 1

GR foci behave as phase-separated condensates. U2OS cells expressing GFP-GR were incubated with Dex and imaged by confocal microscopy. a Representative images of cells incubated with vehicle (Veh) or Dex (Scale bar: 5 μm). The zoomed image corresponds to the region indicated by the dashed square (Scale bar: 1 μm). The white arrows point to some GR foci. See also Additional file 1: Supplementary Video S1. b Representative images of the same cells before and after a treatment with 1% v/v 1,7-heptanediol (1,7-HD) for 30 s (Scale bar: 5 μm). The zoomed images correspond to the regions indicated by the dashed squares (Scale bar: 2 μm). This experiment was run in 20 cells obtaining similar results. c Representative results obtained in FRAP experiments in which the intensity recovery in adjacent regions corresponding to a GR focus (blue circle) and the nucleoplasm (red circle) were analyzed (Scale bar: 5 μm). The zoomed image corresponds to the region indicated by the dashed square (Scale bar: 1 μm). Fluorescence intensity was doubly normalized to the intensity of the same region before bleaching and to the intensity of a non-bleached reference region at the same time point. Time = 0 s corresponds to the beginning of the image sequence showing the fluorescence recovery. Data was fitted considering a simple exponential model (Eq. 2). Fits are shown with a continuous blue (foci) and red (nucleoplasm) lines. Raw data can be found in Additional file 13: Supplementary Table S1. d Time-lapse imaging showing a fusion event between two GR foci, indicated with white arrows before fusion (Scale bar: 5 μm). Fluorescence intensity profiles along the foci at different time points of the image stack. The intensity is represented relative to that determined in the nucleoplasm. Five fusion events were detected in 48 time-lapse sequences of different cells. See also Additional file 5: Supplementary Video S2

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