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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: J Immunol. 2013 May 3;190(11):5506–5515. doi: 10.4049/jimmunol.1300199

FIGURE 1.

FIGURE 1

Expression of GARP and latent TGF-β1 by Treg. (A) GARP and LAP staining of freshly isolated lymph node cells gated on CD4+Foxp3+ and CD4+Foxp3− or CD4-enriched cells stimulated for 24, 48, or 72 h with plate-bound anti-CD3+IL-2. Quadrants are set based on background staining from isotype controls throughout. (B) Sorted CD4+Foxp3-GFP+ cells were immediately stained for GARP (Fresh) or cultured in the presence of anti-IL-2 alone or anti-IL-2 plus IL-2 (human), IL-4, IL-7, IL-9, IL-15, IL-21, or TNF-α, and GARP was measured by FACS. (C) IL-2/anti–IL-2 mAb complexes were injected I.P. on days 1–3. Splenocytes were stained for GARP/LAP on day 5. Gated on CD4+Foxp3+ cells. (D) Freshly isolated lymph node cells gated on CD4+Foxp3+ cells stained with isotype controls, surface stained for GARP/LAP, stained for GARP/LAP before and after fixation/permeabilization, or blocking of GARP/LAP with 10x unlabeled antibody prior to fixation/permeabilization, followed by intracellular staining for GARP/LAP.