Figure S4. Direct target engagement in primary immune cells. (A-B) CD8+ T-cells were purified from spleens, preincubated with inhibitors AZD8835 (α/δ) AZD8186 (β/δ) and CAL101 (δ) for 1 h, then stimulated with 10 μg/mL α-CD3 and 2 μg/mL soluble α-CD28 for 25 min at 37 °C. (A-B) Line graph shows MFI of pAkt(Ser473) and pS6(Ser240/244), histograms show representative data. Data are representative of ≥2 independent experiments. (C) Cell lysates were prepared, separated by SDS-PAGE, and immunoblotted to detect pS6(Ser240/244), pNDRG-1(Thr346) and Vinculin. (PDF 307 kb)