Extended Data Fig. 8: BFAR restrains CD8+ T cell function through USP39.
From: Age-related decline in CD8+ tissue resident memory T cells compromises antitumor immunity
a,b, Immunoblot analysis of the interaction of BFAR with USP7 or USP9X in HEK293T cells transfected with the indicated expression vectors. c, Immunoblot analysis of the interaction of JAK2 with USP9X in HEK293T cells transfected with the indicated expression vectors. d, Ubiquitination of JAK2 in HEK293T cells transfected with the expression vector encoding USP9X, Flag-JAK2 and HA-Ub as indicated. e,f, Ubiquitination of USP9X and USP39 in HEK293T cells transfected with the expression vector encoding Flag-USP9X, Flag-USP39, HA-BFAR and Ub as indicated. g-l, qPCR analysis of the mRNA expression of Bfar and Usp39, immunoblot of phosphorylated (p-) and total STAT1 and flow cytometric analysis of the proliferation and the frequencies of IFNγ-producing CD8+ T cells of WT and BFAR-deficient mouse splenic CD8+ T cells that were reconstituted with EV (control) or LMP-shUSP39 vector and stimulated by plate-bound anti-CD3 plus anti-CD28 (α-CD3/28; 1μg/ml) for 3 days. Bar graphs are presented as mean ± s.e.m. A two-tailed Student’s t-test was performed for comparisons. The data are representative of three independent experiments.
