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. 2014 Nov 5;289(51):35561–35569. doi: 10.1074/jbc.M114.608745

FIGURE 5.

FIGURE 5.

Expression of T-bet in resting naive, effector, TEM, and TCM CD4+ T cells. PBMCs with or without stimulation were stained with surface marker mAbs including anti-CD3, anti-CD4, anti-CD45RA, anti-CD62L, and anti-CCR7 for 30 min at 4 °C, and fixed in fixation buffer for 10 min at 37 °C, permeabilized in ice-cold 90% methanol for 30 min, and labeled with anti-IFN-γ, anti-T-bet (open histogram), or anti-IgG1κ (shaded histogram) mAbs for 50 min at 4 °C. Gated on CD3+ CD4+ T cells, expression of T-bet in resting naive, effector, TEM, and TCM CD4+ T cells distinguished by surface markers CD45RA and CD62L was analyzed (a). The same results were observed based on the surface markers CCR7 and CD45RA (b). Gated on T-bet+ or T-bet− CD4+ T cells, expression of CD45RA, CD62L, and CCR7 was analyzed (c and d). Statistical data are shown for the results from ten independent experiments.