GCRα cellular translocation in human CD4⁺ and CD8⁺ lymphocytes in response to 10⁻⁷ M DEX treatment in vitro. Freshly isolated PBMCs were stimulated for 1 hour with 10⁻⁷ M DEX or remained untreated. Representative images of GCRα cellular translocation in (A) CD4⁺ and (B) CD8⁺ T cells in response to DEX treatment are shown (original magnification × 630; blue, DAPI, nuclear staining; red, cy3, GCRα; green, APC, CD4⁺ or CD8⁺ T-cell surface staining) (the cells from 4 different donors were evaluated in this assay). Note cytoplasmic localization of GCRα before DEX treatment and increase in nuclear localization of the GCRα after DEX treatment in both CD4⁺ and CD8⁺ T cells. See “Materials and methods, GCRα nuclear translocation” for more detailed image acquisition information. (C) GCRα (Cy3) MFI for the nuclear region of CD4⁺ and CD8⁺ before and after DEX treatment. (D) Addition of DEX resulted in significant increase in the amount of nuclear GCR in both purified CD4⁺ and CD8⁺ cells. GCR was measured in nuclear extracts from CD4⁺ vs CD8⁺ T cells plus or minus DEX (1 hour) on the basis of its interaction with GRE consensus motive immobilized to the plate (TransAM GR transcription factor assay). Data are expressed as mean plus or minus SEM (n = 6).