Phenotype of GAD TMr-treated anergic T cells. (A-D) Ann-V^- high- and low-avidity T cells were sorted after TMr treatment and analyzed for CD25 and CTLA-4 expression by flow cytometry. Profiles of control and GAD TMr-treated T cells are shown in gray and black, respectively. (E) Western blot of FoxP3 expression in high- and low-avidity T cells. The membrane was probed with an anti-FoxP3 antiserum (top) and subsequently with an anti-TFIIB Ab (bottom) to verify equal loading of lanes. The band obtained from FoxP3-transfected 293T cells is shown as control in the last lane. Bar graph shows densitometric analysis of FoxP3 bands normalized to TFIIB bands detected in each lane.