Research Paper Volume 5, Issue 1 pp 18—36

Figure 1. NF-κB p65/RelA nuclear expression in human peripheral blood CD4⁺ T lymphocytes activated via the T cell antigen receptor.

Purified CD4⁺ T cells were incubated at 37°C with plate-bound anti-CD3 for 2h or 4h; control cells were incubated for 4h without additional treatment. (A) Nuclear extracts (NE) were fractionated by SDS-PAGE, transferred to PVDF membranes and probed with antibodies directed against p65/RelA, transcription factors SP1 and TATA binding protein (TBP), or β actin. p65/RelA levels from 3 donors were compared to varying concentrations of a control cytoplasmic extract (CE). Induced p65/RelA levels in all 29 donors are shown in Table S3. (B) Fold-increase of nuclear p65/RelA treated cells was calculated relative to the levels in untreated cells (Table S4). The average fold increase of nuclear p65/RelA for 29 donors after 2h or 4h activation separated by subjects less than 65 years (Y) and 65 and over (O) is shown in the graph. p-values shown above bar graphs compare the fold induction between 2h and 4h of activation in Y and O groups, and were determined by Student's test (p<0.05 was considered as significant). Error bars reflect the standard error of the mean (±SEM) (in young 0.41; 0.52) (in old 0.38; 0.39). (C) Fold-increase of nuclear p65/RelA treated cells was calculated relative to the levels in untreated cells (Table S4). The average fold increase of nuclear p65/RelA for 21 donors after 2h or 4h activation separated by subjects 50-64 years (M) and 65 and over (O) is shown in the graph. p-values shown above bar graphs compare the fold induction between 2h and 4h of activation in M and O groups, and were determined by Student's test (p<0.05 was considered as significant). Error bars reflect the standard error of the mean (±SEM) (in middle 0.66; 0.83) (in old 0.38; 0.39).