Key signaling pathways for IL-2 transcription are activated in TCR/CD3 stimulated naive CD4⁺ T cells of CB.

Abstract

<p>(A) Protein expression by Western blot of ERK1/2 and phosphorylated ERK1/2Tyr202/ Tyr204 (pERK1/2) in naive CD4⁺ T cells of CB as well as for naive CD31⁺ adult T cells. The ratios of relative protein expression levels are indicated below the respective bands. Data are representative of two independent experiments. (B) Whole cell protein extract of NFATc2 and phosphorylated NFATc2 (pNFATc2) in CB naive CD4⁺ T cells and adult naive CD31⁺ T cells under different stimulation conditions. The densitometric analyses of the immunoblot detection for the relative protein expression level are shown as ratio of NFATc2 or pNFATc2 to αTubulin. Lysates from three different donors were pooled. Data are representative of at least three independent experiments. αTubulin was used as a loading control. (C-D). The NFATc2 protein expression in cytoplasm or nucleoplasm (separated through a dashed line) in (C) naive CD4⁺ T cells of CB or (D) CD31⁺ naive T cells of adult were detected and the phosphorylated (pNFATc2) and dephosphorylated (NFATc2) forms quantified. Cells were stimulated as indicated in the presence or absence of cyclosporin A (CsA). One representative experiment out of two comparable experiments is shown. unstim. = unstimulated.</p