PBMC cells were separated from human blood from healthy volunteer donors following existing protocol 82 and pelleted by centrifugation at 1000 g for 2 min. The supernatant was discarded and the PBMC pellet was resuspended in PBS containing 2% FBS for washing. The suspension was centrifuged at 1000 g for 2min again and the PBMC pellet was resuspended in RPMI-1640 (Gibco) media supplemented with 10% FBS. The cells were then seeded into 6-well plates and treated with testing compounds at indicated concentrations and time period. After treatment, the cells were collected into 2-ml eppendorf tube and centrifuged at 500g for 2 min to pellet the cells, the supernatant was discarded, and the pellet was resuspended in 1 ml PBS and centrifuged at 500 g for 2min again. The PBMC pellet was lysed with 60 µL of lysis buffer containing 50 mM Tris-HCl, pH 7.5, 1% (v/v) Triton X-100, 1 mM EGTA, sodium orthovanadate, 50 mM NaF, 0.1% (v/v) 2-mercaptoethanol, 10 mM 2-glycerophosphate, 5 mM sodium pyrophosphate, 0.1 µg/ml mycrocystin-LR (Enzo Life Sciences), 270 mM sucrose, 0.5 mM DIFP (Sigma, Cat# D0879) in addition to complete EDTA-free protease inhibitor cocktail (Sigma-Aldrich Cat # 11836170001). DIFP is highly toxic and must be prepared in a fume hood to a stock solution of 0.5 M in isopropanol. The lysed cells were then centrifuged at 1500 g for 15 min at 0 °C. The supernatants were collected for analysis by quantitative immunoblotting. For long term storage, the supernatant was flash frozen and stored at -80°C. Protein concentrations of cell lysates were determined using Pierce™ BCA Protein Assay Kit (ThermoFisher).