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

Figure 4.

Figure 4

MyD88 deficiency inhibited IL-36β-mediated mTORC1 activation of CD8+ T cells and resulted in suppression of CD8+ T function. Naïve CD8+ T cells were, respectively, isolated from C57BL/6j and MyD88−/− mice and stimulated with or without plate-bound 10 μg/ml anti-CD3 mAb, in the presence or absence of IL-36β (100 ng/ml) for various lengths of time. (A) Degradation of IκB at 1h upon stimulation was measured by western blot. (B) Phosphorylation of ribosomal protein S6 at 48 h was measured and then analyzed by western blot. (C) Phosphorylation of ribosomal protein S6 at 48 h was measured and then analyzed by flow cytometry. (D) The levels of IL-2 and IFN-γ production in the supernatants were measured by ELISA method at 48 h. ***p < 0.001 by unpaired t-test. (E) Cell sizes (forward scatter) at 72 h were determined by flow cytometry. (F) 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.