Extended Data Fig. 8: The TXA2 receptor TP regulates CD8+ T cells in an ARHGEF1-dependent manner.
From: Aspirin prevents metastasis by limiting platelet TXA2 suppression of T cell immunity
a-b, Proliferation WT and Arhgef1-deficient naïve CD8+ T cells electroporated with nucleoprotein complexes of Cas9 and 3 different sgRNAs targeting Tbxa2r or scrambled sgRNA control and stimulated in vitro with anti-CD3/28 antibodies and rhIL-2 in the presence or absence of 5 μM TXA2 analogue U46619. Proliferation is assessed as dilution of the CellTrace Violet. n = 5 independent replicates per condition. c, Confirmation of Tbxa2r deletion using SDS-PAGE and Western blotting of cell lysates from cells in (a, b). d, Number of cells 5 days after stimulation of naïve FACS-sorted CD8+ T cells with anti-CD3/28 antibodies and rhIL-2 in the presence of TXA2 analogue U46619 or vehicle control and the treatment with TP inhibitor SQ 29548 (10 μM). n = 4 independent replicates per condition. Data are representative of two independent experiments (a-d). Multiple two-tailed Student’s t-test with Holm-Šídák correction (b) and 2-way ANOVA with Tukey multiple comparisons test (d). Graphs show mean ± s.e.m.
