Fig. 3.

Differentiation between activated and regulatory CD4⁺ CD25⁺ SkIL and LNC: (a) SkIL and (b) draining LNC of healthy, AA and SADBE-treated AA mice were stained with anti-CD25, -CD4, -CD152 and -CD154 and were triple-stained with anti-CD4 (FITC or APC), anti-CD25 (PE) and either anti-CD152 (FITC) or anti-CD154 (APC). The 1st and 2nd rows show single-staining overlays for CD25, CD4, CD152 and CD154. The 3rd, 4th, 5th and 6th rows show negative controls and double-staining with anti-CD25/anti-CD152 or anti-CD154 and anti-CD4/anti-CD152 or anti-CD154. In the 7th row the population of CD4⁺ CD25⁺ cells has been gated. It shows all CD152⁺ and all CD154⁺ cells, but only CD4⁺ CD25⁺ double-positive cells (as defined by the gating). Thus, cells in the upper right quadrant are CD4⁺ CD25⁺ CD152⁺, respectively, CD4⁺ CD25⁺ CD154⁺.