Fig. 1: Molecular compound screening identifies JQ1 that suppresses PD-1 expression of Jurkat cell and T cell.

A Flow diagram of the experimental design. B Plot of the ratio of PD-1 expression on Jurkat cells treated with the indicated compounds as detailed in Supplemental Table 5 (n = 3). C Plot of the ratio of PD-1 expression on Jurkat cells stimulated with 150 ng/mL PHA and the same compounds as that shown in Supplemental Table 5 (n = 3). D Jurkat cells were treated with 0.5 μM JQ1 with and without PHA (150 ng/mL) stimulation, and PD-1 expression was determined by FACS at the indicated time points (n = 4). E Jurkat cells were treated with 0.5 μM JQ1 with and without PHA (150 ng/mL) stimulation, and PD-1 expression was determined by qRT-PCR at the indicated time points (n = 3). F Jurkat cells were treated with indicated doses of JQ1 for 24 h in the presence or absence of PHA (150 ng/mL), PD-1 expression was determined by qRT-PCR (n = 5). G Jurkat cells were treated with indicated doses of JQ1 for 24 h in the presence or absence of PHA (150 ng/mL), PD-1 expression was determined by FACS (n = 4). H CD8 + T cells from healthy donors were treated with 0.5 μM JQ1 for 24 h with and without P/I (PMA: 20 ng/L, ionomycin: 0.5 μM) stimulation, and PD-1 expression was determined by FACS (n = 3). I CD4 + T cells from healthy donors were treated with 0.5 μM JQ1 for 24 h in the presence and absence of PHA (500 ng/ml), P/I (PMA: 20 ng/L, ionomycin: 0.5 μM), or anti-CD3/CD28 (anti-CD3: 200 ng/ml, anti-CD28: 200 ng/ml), and PD-1 expression was determined by FACS (n = 4). J CD8 + T cells from healthy donors were treated with 0.5 μM JQ1 for 24 h with and without P/I (PMA: 20 ng/L, ionomycin: 0.5 μM) stimulation, and CD69 expression was determined by FACS (n = 3). K CD4 + T cells and CD8 + T cells were treated with 0.5 μM JQ1 for 24 h, and the secretions of IL-2 were evaluated by ELISA (n = 4). L CD4 + T cells and CD8 + T cells were treated with 0.5 μM JQ1 for 24 h, and the secretions of IFN-γ were evaluated by ELISA (n = 4). M CD4 + T cells and CD8 + T cells were treated with 0.5 μM JQ1 for 24 h, and the secretions of TNF-α were evaluated by ELISA (n = 4). Data in (B–M) were expressed as mean ± SD. n = 3 or more independent biological replicates, presented as individual points. P value < 0.05 was considered to be significant (D–G: two-way ANOVA, B, C, H–M: one-way ANOVA with Bonferroni post hoc test).