Skip to main content
Fig. 4 | Journal for ImmunoTherapy of Cancer

Fig. 4

From: 1-Pyrroline-5-carboxylate released by prostate Cancer cell inhibit T cell proliferation and function by targeting SHP1/cytochrome c oxidoreductase/ROS Axis

Fig. 4

SHP1 Inhibition Weaken the Effect of PCC Metabolites on T Cells. (a) Human primary CD3+ T cells were pretreated with PCM and the inhibitor of SHP1 NSC87877 (12.5 μM) then stimulated for 3 days with anti-CD3/CD28 beads. Shown is the percentage of cell proliferation by flow cytometry. The right side of bar graph is the representative result by flow cytometry. (b, c) Human primary CD3+ T cells were pretreated with PCM and the inhibitor of SHP1 NSC87877 (12.5 μM) then stimulated for 3 days with anti-CD3/CD28 beads. Shown is the levels of total and mitochondria ROS by flow cytometry. The right side of bar graph is the representative result by flow cytometry. (d) Jurkat cells treated with PCM and the inhibitor of SHP1 NSC87877 for 24 h. Shown is the percentage of cell proliferation by CCK-8 assay. (e, f) Jurkat cells treated with PCM and the inhibitor of SHP1 NSC87877 for 24 h. Shown is the percentage of cell proliferation by CCK-8 assay. The right side of bar graph is the representative result by flow cytometry. (g-j) Human primary CD3+ T cells were pretreated with PCM and the inhibitor of SHP1 NSC87877 (12.5 μM) then stimulated for 3 days with anti-CD3/CD28 beads.. Supernatants from cell cultures were analyzed for cytokines levels using commercially available ELISA kits, including IL-2, TNF-α, IL-6, IL-17A. (k) Jurkat cells treated with PCM and the inhibitor of SHP1 NSC87877 for 24 h. Shown is the relative activity of CIII by microplate reader. All experiments were repeated at least three times. Error bars are SEM of biological replicates and **p < 0.01; ***p < 0.001

Back to article page