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. 2019 Aug 7;10:1803. doi: 10.3389/fimmu.2019.01803

Figure 2.

Figure 2

IL-36β-mediated CD8+ T cell activation was dependent on mTORC1. Naïve CD8+ T cells were isolated from C57BL/6j mice and stimulated with plate-bound 10 μg/ml anti-CD3 mAb, in the presence or absence of IL-36β (100 ng/ml), or rapamycin (20 or 50 nM) for various lengths of time. (A) Phosphorylation of ribosomal protein (p-S6) was measured by flow cytometry at 48 h. (B) Phosphorylation of ribosomal protein (p-S6) was measured by western blot at 48 h. (C) The levels of IL-2 and IFN-γ production in the supernatants were measured by ELISA method at 48 h. (D) Cell sizes (forward scatter) at 72 h were determined by flow cytometry. (E) Cell proliferation based on CFSE dilution assay at 72 h were determined by flow cytometry. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01 and ***p < 0.001 by unpaired t test. The experiment was repeated independently three times.