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. Author manuscript; available in PMC: 2014 Jan 9.
Published in final edited form as: J Immunol. 2010 May 7;184(12):10.4049/jimmunol.0901479. doi: 10.4049/jimmunol.0901479

FIGURE 4.

FIGURE 4

Nod2 mRNA is expressed in human Treg cells. A, RNA extracted from freshly isolated CD4+CD25++ Treg cells, CD4− PBMCs, and 293T cells is probed for Nod2, CD11b, and actin. Expression of Nod2 mRNA within Treg cells and CD4− PBMCs (CD4−) is significantly increased over 293T cells (mean, 9.34 ± 1.30, 29.73 ± 3.55; p = 0.0077). The inset DNA gel demonstrates an absence of the CD11b mRNA transcript in Treg cells, evidence against significant cell subset contamination. Six unique healthy blood donors were used for these experiments, and results represent the mean/SE of each independent experiment. B, The CD4+CD25++ human T cell population is highly purified. The upper flow cytometry dot plots demonstrate >98% purity of isolated cells for CD4 and 78% purity for FOXP3. The bar graph demonstrates the mean (SE) of CD4/FOXP3 enrichment for the six unique blood donors. C, RNA extracted from EGFP+FOXP3+ murine splenocytes or CD4− murine splenocytes demonstrates significant upregulation of murine Nod2 mRNA transcripts compared with murine muscle negative control (21.35 ± 0.48, p < 0.0005; 15.58 ± 0.58, p = 0.016, respectively).