Extended Data Fig. 6: TCPOBOP promotes CAR-dependent gene expression in ex vivo-isolated effector T cells.
From: CAR directs T cell adaptation to bile acids in the small intestine
a, Top left, equal numbers of CD45.1 wild-type (B6; blue) and CD45.2 CAR-deficient (B6.Nr1i3−/−; red) naive CD4+ T cells were transferred together into B6.Rag1−/− mice. The resulting Teff cells were FACS-purified from spleen after 3 weeks. Right, sequential gating strategy for re-isolating wild-type and CD45.2 CAR-deficient spleen Teff cells. Bottom left, mean ± s.e.m. relative expression of Abcb1a (n = 4), Cyp2b10 (n = 4), and Il10 (n = 3), determined by qPCR, in ex vivo-isolated wild-type (B6) or CAR-deficient (B6.Nr1i3−/−) spleen Teff cells. These cells were used for ex vivo stimulation experiments with or without small-molecule ligands (b, c). P values (paired two-tailed Student’s t-test) are indicated. b, Mean ± s.e.m. relative expression of Abcb1a (n = 4), Cyp2b10 (n = 4), and Il10 (n = 3), by qPCR, in wild-type (B6) or CAR-deficient Teff cells isolated from transferred Rag1−/− mice (as in a), and stimulated ex vivo with anti-CD3 and anti-CD28 antibodies in the presence or absence of TC (10 μM), the mCAR inverse agonist And (10 μM), or both. P values (one-way ANOVA with Tukey’s correction for multiple comparisons) are indicated; ****P < 0.0001. c, Mean ± s.e.m. relative expression of Abcb1a, Cyp2b10, and Il10 (n = 5), determined by qPCR, in wild-type (B6) or CAR-deficient (B6.Nr1i3−/−) Teff cells isolated and stimulated as in a, b in the presence or absence of TC (10 μM) or PCN (10 μM). Data are presented as fold change in mRNA abundance relative to vehicle-treated cells (DMSO for TC; ethanol for PCN). ****P < 0.0001, one-way ANOVA with Dunnett’s correction for multiple comparisons. NS, not significant.
