Figure 1. Kinetics of GRα and GRγ activation.

(A) Protein Workshop was used to illustrate conformational differences of GR isoforms bound to FKBP5 promoter (PDB ID 3G6U and 3G6T⁸). GRγ (blue) differs from GRα (yellow) by a single arginine residue inserted in the DBD, which is a DNA and protein interaction surface. The additional arginine in the DBD is highlighted in red, with adjacent amino acids shown in green. (B) Protein Workshop was used to illustrate the proximity of the additional arginine to NLS1, shown in white. (C) Cells were co-transfected with pG-EGFP GRα and pG-dsRedXP GRγ, cultured in charcoal stripped serum, treated with 100 nM dexamethasone and imaged using time-lapse microscopy. An example of GRα (green) and GRγ (red) expression in the same cell is shown. Nuclear translocation from multiples cells were quantified. Graphs (mean +/− SD) are representative of three independent, and a total of 71 cells were analysed. Samples were compared with an unpaired, two-tailed Student’s t test (*P < 0.05). (D) Cells were transfected with pG-dsRedXP GRγ, cultured in charcoal stripped serum and imaged using time-lapse microscopy. Expanded regions are highlighted with white boxes. Arrows indicate GRγ at membrane ruffles.