Figure 5

SHP-2 activation by fusaruside suppresses p-STAT1/T-bet and production of Th1 cytokines upon TCR stimulation. Total TIL were stimulated with anti-CD3/-CD28/hIgG1 beads (bead: cell = 10:1) or anti-CD3/-CD28/PD-L1 beads plus 100 ug/mL anti-PD-1 blockade (BMS-936558) for 48 h in the presence of 50 uM fusaruside or DMSO. Then p-STAT1 and T-bet were analyzed by flow cytometry. Supernatants were collected and stored at -80. Th1 (IFN-γ and IL-2) and Th2 (IL-10) cytokines in the supernatants were determined by Luminex. A) Summary data of frequency of p-STAT1+ and T-bet+ cells in CD8⁺ and CD4⁺ TIL at different conditions is shown (n = 6). B) Summary data of amount of IFN-γ (n = 8), IL-2 (n = 4) and IL-10 (n = 8) in the supernatants of TIL cultured under indicated conditions. The graphs present the mean±SEM from different HNC patients. Statistical significance was determined by Wilcoxon (non-parametric paired) test. *p < 0.05, **p < 0.01.