Figure 3

ERK in CD8⁺ T-cells was phosphorylated by CD3/28 stimulation and inhibited by trametinib. Pre-expanded CD8⁺ T-cells from HD6 were supplemented with 100 IU/mL IL-2, with or without 10 ng/mL TGF-β1, and a kinase inhibitor for 3 h. CD8⁺ T-cells were restimulated with CD3/28 beads. Experimental methods are shown in (a). CCR4 expression was evaluated using flow cytometry on day 2 (b). The same samples as in (b) were collected 10 min after stimulation, and phosphorylation of ERK and STAT3 was evaluated by western blotting (c). In the image in (c), the left-most and second left-most lanes are transposed. Original images are shown in Supplementary Fig. S7. t total, p phosphorylated.