Differential expression of galectin-10 protein and mRNA in CD4⁺CD25⁻ T cells and CD25⁺ Treg cells. (A) Selected gel regions showing the different protein spot intensities of galectin-10 isoforms A-C in resting and activated human CD4⁺ T cells and CD25⁺ Tregs. Due to gel size, the IEF rod gels were cut in 2 halves at approximately pI 6.0 to 6.4 before further processing. Arrowheads show cutting edges, which were due to technical reasons. In this context, please also see Figure S6. (B) The ratios of protein spot intensities were calculated on the basis of 2D PAGE from sets of T cells derived from 4 individual human volunteers (performed as triplicates each; mean plus or minus SD from each triplicate shown; asterisks indicate P values as defined in “Materials and methods, Statistical analysis”). For proteome analysis, freshly isolated (leukapheresis products) as well as stimulated (leukapheresis products and buffy coats) T-cell populations were used. Due to limited recovery of CD25⁺ Treg cells isolated from buffy coats, only stimulated T-cell populations were analyzed. (C) Quantification of relative galectin-10 (Gal-10) mRNA levels compared with Foxp3 in freshly isolated CD4⁺ T cells and CD25⁺ Treg cells. cDNA samples were subjected to qRT-PCR. The relative quantities of galectin-10 and Foxp3 mRNA were normalized according to the expression of EF1-α mRNA. The qRT-PCR data are representative of 3 independent experiments; measurements were performed in triplicates; mean plus or minus SD from each triplicate shown; asterisks indicate P values as defined in “Materials and methods; Statistical analysis”.