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. 2021 Aug 20;9(8):e002279. doi: 10.1136/jitc-2020-002279

Figure 3.

Figure 3

ZSTK474 inhibits PI3K signaling not only in Tregs but also in CD8+ T cells. (A, B) Phosphorylation status of SLP-76 at Y128 (p-SLP-76) and Akt at S473 (p-Akt) after stimulation with anti-CD3/anti-CD28 mAb at the indicated time points with the indicated doses of ZSTK474. Representative flow cytometry histograms (A) and summaries of mean fluorescence (MFI) of p-SLP-76 and p-Akt (B) in Tregs, helper CD4+ T cells and CD8+ T cells. Data are means±SE. (C) Phosphorylation of Akt at S473 (p-Akt) and S6 at S235/236 (p–S6) after stimulation with anti-CD3/anti-CD28 mAb for 20 min with or without ZSTK474 (1 or 10 µM). Representative pictures of immunoblot analyses (left) and the average of quantification values of bands (right). Data are means±SD. Statistical analyses were performed by Student’s t-test (B), and Dunnett’s test (C). *P<0.05; **p<0.01; ***p<0.001. mAb, monoclonal antibody.