p110δ regulates TCR signaling by human T cells. (A) T cells from healthy human donors were pretreated with DMSO alone, IC, or LY and stimulated with anti-CD3 and anti-CD28 for 5 minutes. Lysates were immunoblotted to examine the levels of total and phosphorylated Akt and Erk. Data represent one of 3 donors. (B-F) T cells from healthy human donors were pretreated with DMSO alone, IC, or LY and stimulated with anti-CD3 and anti-CD28 for different time points. The pooled bar-coded aliquots were stained for surface markers and phospho-proteins and analyzed by flow cytometry. Populations were gated into naive (N) and effector/memory (E/M) CD4⁺ and CD8 ⁺ T cells, and changes in phosphorylation were assessed for each population. Histograms showing dynamics of phosphorylation of Akt (B), GSK3β (C), Erk (D), and S6 (E) in DMSO-treated samples are shown. (F) The data for DMSO- and inhibitor-treated samples were converted into a heatmap showing fold change compared with unstimulated control. The data presented are representative of 3 independent experiments with different donors.