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. 2012 Aug 17;61(9):2340–2348. doi: 10.2337/db12-0049

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© 2012 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 4. — Rapamycin/IL-2 combination therapy modulates pAkt, but not phosphorylated (p)Erk or pS6, in CD4⁺ T cells. Thawed PBMCs were stimulated with media alone or cross-linking of anti-CD3/anti-CD28–bound antibodies with Fab x-linker for 20, 30, or 10 min for Erk, S6, and Akt analysis, respectively. Cells were then fixed, permeabilized, and stained for CD4, CD8, pAkt, pErk, and pS6. Representative histograms are shown for pErk (A), pS6 (B), and pAkt (C) with media (gray line) and anti-CD3/anti-CD28 (black line) stimulation. Fold increase (FI) was determined by the equation (MFI with CD3/CD28 stim) ÷ (MFI with media) for each sample and is compared between day 0 and 84 for pErk (A), pS6 (B), and pAkt (C). Data from all subjects receiving full dosing of rapamycin are included except for pAkt for subjects 1 and 2, for which pAkt data were not acquired. Statistical significance was determined by a paired Student t test.