Actions of 8-NH₂-Ado on Akt and Erk signaling pathways of peripheral blood monocytes. Freshly isolated PBMCs in 10% fetal bovine serum (FBS), RPMI were incubated continuously with 3μM 8-NH₂-Ado for 0, 5, and 17 hours. Lymphoma cells from a lymphoma patient (MCL) were collected from the peripheral blood and allowed to proliferate in culture (20% FBS in Dulbecco modified Eagle medium). Once sufficient cell counts were obtained, the lymphoma cells were incubated continuously with 3μM 8-NH₂-Ado for 0, 5, and 17 hours. JeKo cell lysates (Figure 4) were used for comparison. Cell lysates (30 μg) were immunoblotted for phospho- and total protein levels as shown. GAPDH was used as a loading control.