NFAT2 is overexpressed and to some extent localized into the nucleus of CLL B cells. (A) Relative NFAT2 transcript levels were assessed by quantitative RT-PCR from freshly isolated normal blood peripheral B and T lymphocytes (normal lymphocytes B and T; n = 6 and n = 3, respectively) and from CLL B cells from NR and R cases (n = 17). NFAT levels were normalized to Abelson expression. (B) Freshly purified CLL B cells (n = 10) were immunostained with anti-NFAT2 antibody. Nuclei were counterstained with 4,6-diamidino-2-phenylindole. One representative case of each CLL group (NR and R) is presented. Images were acquired using confocal laser-scanning microscope (TCS SP2; Leica). Scale bars represent 5 μm. Single optical sections were obtained with high numerical aperture lens (63 × 2.8 NA) to determine the percentage of NFAT2-positive nuclei in NR and R cells and graphed on the right histogram; an average of 90 cells/sample (6 samples) were analyzed. No statistical difference was observed (ns).