Figure 6.

DRD5 does not form heteromeric complexes with CXCR4. (A) Jurkat cells were transfected with constant amount of cDNA codifying for RLuc-fused to DRD5 (as donor) and increasing amounts of cDNA encoding for the YFP fused to CXCR4 (as acceptor). The relative amount of BRET was quantified as the ratio between YFP fluorescence and RLuc activity (×1000) and expressed as mBU. Each symbol represents data obtained from an individual determination. Data are from 3 independent experiments. Best fit of data indicates a linear relationship (the equation is indicated), suggesting the absence of physical interaction. (B) Naïve CD4⁺ T cells (CD3⁺ CD4⁺ CD45RB^high) were isolated from the spleen of wild-type mice and then activated with anti-CD3/anti-CD28 mAbs-coated Dynabeads in the presence of IL-2 and RA for 5 days to induce gut tropism. Afterward, cells were incubated with TM7C (4 μM) or TM6C (4 μM) peptides for 4 hours. Cell migration to CXCL12 (300 ng/mL) was determined in transwell assays after 3 hours. Each symbol represents data obtained from an individual determination (n = 4–7 mice per group). Mean ± SD are indicated. ∗∗∗P < .001; ∗∗∗∗P < .0001 by 2-way ANOVA followed by Sidak’s post hoc test.