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. Author manuscript; available in PMC: 2009 Jul 15.
Published in final edited form as: J Immunol. 2008 Jul 15;181(2):1272–1281. doi: 10.4049/jimmunol.181.2.1272

FIGURE 3.

FIGURE 3

Enhanced basal c-Rel expression in Pre-Ac CD4+ T cells and inhibition by PF. A, Pre-Ac (blue) or naïve (green) WT CD4+ T cells were stimulated with α-TCR-β + α-CD28 for the times indicated, paraformaldehyde fixed, stained with α-c-Rel, α-CD4, and DRAQ5, and then analyzed for intracellular c-Rel expression and c-Rel nuclear translocation using the ImageStream® 100 multispectral imaging flow cytometer. B, ChIP. SYBR® green qPCR, and primer set C [−201 to −111] were used to determine c-Rel promoter occupancy in naïve and Pre-Ac WT CD4+ T cells following α-TCR-β + α-CD28 for the times indicated; data are expressed as fold induction relative to unstimulated naïve cells. C, Pre-Ac (blue) or naïve (green) IL-2-GFPKI/WT CD4+ T cells were stimulated with α-TCR-β + α-CD28; PF or CsA was added at various times after T cell activation as indicated. Cells were harvested at 54 h (naïve) or 24 h (Pre-Ac) and gated on 7-AAD−CD4+ T cells for GFP analyses. D, PF or CsA were added to IL-2-GFPKI/WT CD4+at the time of α-TCR-β + α-CD28 stimulation. Cells were harvested for GFP analyses at the times indicated.