Extended Data Fig. 2. NUR77 restrains survival and proliferation of BCR-stimulated B cells in a cell-intrinsic manner.
A. Graph depicts live CD45.2+ B cell number corresponding to Fig. 2A, B samples. B. Lymphocytes from NUR77-EGFP mice were stimulated with anti-IgM +/− 20 ng/mL BAFF for 24 h. Graph depicts GFP MFI of live B cells. C, D. Graphs depict the ratio of CD45.2+ Nr4a1−/− or Nr4a1+/+ B cells relative to co-cultured WT CD45.1+ B cells, normalized to the input ratio, and correspond to Fig 2. C, D samples. E. Splenocytes from Nr4a1+/+, Nr4a1+/− and Nr4a1−/− mice were stimulated with 10 μg/mL anti-IgM for 2 h. Graph depicts MFI of endogenous NUR77 expression in CD23+ B cells. F-H. Either MACS-purified B cells or total lymphocytes from CD45.2+ Nr4a1−/− and CD45.1+ Nr4a1+/+ mice were co-cultured in a 1:1 ratio with anti-IgM for 72 h. F. Histograms depict CTV dilution after culture with 6.4μg/ml anti-IgM. G. Graph depicts ratio of Nr4a1−/− CD45.2+ relative to co-cultured WT CD45.1+ purified B cells, normalized to the input ratio. H. Graph depicts division index of co-cultured purified B cells. I-J. Lymphocytes from either Nr4a1+/+ or Nr4a1−/− CD45.2+ IgHEL Tg mice were treated as in Fig. 2A. I. Graph depicts ratio of CD45.2+ B cells of each genotype relative to co-cultured CD45.1+ WT B cells, normalized to the input ratio. J. Graph depicts division index of CD45.2+ B cells for each genotype. K, L. Lymphocytes from either mb1-cre, mb1-cre Nr4a1fl/fl, or Nr4a1−/− mice were mixed 1:1 with CD45.1+ lymphocytes and treated as in Fig. 2A. K, L. Graphs depicts total B cell number (K) and division index (L) of each genotype. Data in this figure depict N=3 biological replicates. Mean +/− SEM displayed for all graphs. Statistical significance was assessed with two-tailed unpaired student’s t-test with Holm-Sidak (A-D, G-J); two-way ANOVA with Tukey’s (K, L). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001