Figure 2. Nur77 limits survival of “anergic” IgHEL Tg B cells chronically exposed to cognate HEL in vitro and in vivo Ag.

A-D. Lymph nodes were harvested from IgHEL/sHEL Tg chimeras generated as described in Fig 1C. Tg CD45.2+ B cells were mixed 1:1 with CD45.1+ WT B cells and plated at a total concentration of 5×10⁶ cells per well. These cells were incubated for up to 2 days with or without 20 ng/mL BAFF. The CD45.2/CD45.1 ratio was calculated at each time point and normalized to the CD45.2/CD45.1 cell input ratio. N=4 for each group. E. Schematic for experimental setup used for Fig. 2F-I. IgHEL/sHEL Tg chimeras were generated as in Fig 1C. Splenocytes from IgHEL/sHEL Tg chimeras and CD45.1 WT mice were loaded with the vital dye Cell Trace Violet (CTV) and combined 1:1. 5×10⁶ total splenocytes were adoptively co-transferred into either WT or sHEL Tg mice via I.V. injection. 2 days after adoptive transfer, mice were sacrificed, and spleens were harvested for surface marker analysis via flow cytometry. Donor cells were identified as CTV⁺, and IgHEL Tg and CD45.1 WT B cells were separated based on CD45.1 and CD45.2 expression. F. Representative plots shown. G. Nur77-eGFP MFI of CTV+, CD45.2+ and Nur77-eGFP+ B cells isolated from sHEL Tg+ and sHEL Tg- host mice, N=3 per group. H. IgM^a MFI of adoptively transferred B cells, N=3 per group. I. Ratio of transferred IgHEL Tg cells compared to competitively transferred CD45.1 WT cells normalized to the input ratio. N=3 per group.