Supplementary Figure 5: Reversal of anergy in polyclonal CD4⁺ T cells results in autoantibody production after ablation of newly generated T_reg cells.

Lymphopenic Tcra^−/− mice were given an adoptive transfer (10⁵) of either naive or anergic syngeneic Foxp3^DTR polyclonal CD4⁺ T cells, followed by every other day treatment with diphtheria toxin (DT). Sera were recovered from mice 21 days later and used to probe various tissue extracts (as indicted). (a) Sera taken from Rag^−/− (top), Tcra^−/− (middle), and WT B6 (bottom) mice were assayed as a negative control for autoantibody generation. (b) Sera obtained from three separate adoptive transfer recipients of naive T cells or (c) anergic T cells. Arrowheads indicate self antigen-binding by serum antibodies that are uniquely present with recipients of anergic CD4⁺ T cells. m = marker, He = heart, Ki = kidney, Pa = pancreas, Li = liver, Lu = lung, Gu = gut, Sa = salivary gland. Summary data are shown in figure 5e. Note that an irrelevant 60 Kd background band (most prominent in lung extracts) was demonstrated in all blots even in the absence of serum antibody (not shown), and this band is disregarded.