Figure 1

Phosphorylation of AKT and p38MAPK protein kinases in AKT-S, p38MAPK-S, double signaler (DS) and nonsignaler (NS) categories of CLL samples following stimulation with anti-CD180 mAb. (A and B) CLL cells and control B cells were incubated with anti-CD180 mAb for 20 min (CD180 mAb) or left unstimulated in medium (Medium), washed, stained with anti-CD19 mAb, fixed, permeabilized and stained with anti-p-AKT(Ser473) (A) or with p-anti-p38MAPK (B) mAbs as described in the Materials and Methods, analyzed by flow cytometry and expressed as percentages of positive cells. P values were calculated using the paired t test. (C) Representative immunoblots with the levels of total AKT, p-AKT(Ser473), total p38MAPK and p-p38MAPK in CLL samples following stimulation with anti-CD180 mAb and in unstimulated cultures (Medium) as described in the Materials and Methods. Bcl-2 was used as a loading control. (D and E) Relative optical density, normalized to Bcl-2, for p-AKT and p-p38MAPK immunoblots respectively, n = 6.