FIGURE 2.

Gating strategies used in this study. A, cell surface molecule staining of the live lymphocyte population was gated and confirmed by 7-aminoactinomycin D (7ADD) negative staining. CD4⁺ T cells were then gated for cell surface molecules and/or CD45RO⁺ memory T cell. B, for intracellular staining, cells were first stained for cell surface markers, followed by fixation and permeabilization. The lymphocyte population was then gated, CD4 naive and memory T cells were identified, and various intracellular proteins were examined within the identified CD4⁺, CD45RO⁺ central (CCR7⁺), or effector (CCR7⁻) memory T cells. C, culture of DC, or D, DC co-cultured with isolated CD4⁺ T cells were gated, based on FSC and SSC and these populations were examined for cell surface or intracellular molecules. E, schema of the dual stress-activated pathways, leading to CD4⁺CD45RO⁺ memory T cells, cytokines, and IFN-stimulated genes.