Fig. 7
Expression of A20 and cIAP1 by flow cytometry. MNCs activated with anti-CD3/CD28 for 48 h and, then exposed to TNF-α for 10 min. Cell were first surface stained by CCR7 FITC, CD45RA APC, CD8PerCP antibodies and isotype controls. Stained cells were then fixed and permeabilized, and then stained with purified antibodies to cIAP1and A20 and isotype controls. Cells were washed and incubated with secondary PE conjugated goat anti- rabbit antibody. First cells were gated for CD8+ T cells, and then gated for TN (CD8+, CD45RA+ CCR7+) and TCM T-cells (CD8 + CD45RA- CCR7+) cells. These gated cells were then analyzed for the expression of cIAP1 and A20. a is a representative FACS plot. Blue line represents isotype control, and red line is for A20 and cIAP1. b shows cumulative data for MFI from 5 young and 5 aged subjects. TN and TCM CD8+ T cells from aged subjects show significantly decreased (P < 0.001) expression of both A20 and cIAP