Figure 5.

Modulation of CD180-mediated signaling by sensitization with anti-IgM F(ab)₂. (A–C) Control B cells or CD180⁺sIgM⁺CLL samples were treated with anti-CD180 mAb or first with anti-IgM F(ab)₂ followed by anti-CD180 as described in the Materials and Methods. Unstimulated cultures (Medium) were used as controls. The cells were stained with anti-CD19, fixed, permeabilized and stained with antibodies to p-AKT-P and p-p38MAPK. P values were calculated using the paired t test. (D) Representative immunoblots of p-AKT, p-PI3K, p-p38MAPK, Mcl-1 and Bcl-xL following sequential ligation of sIgM followed by CD180. Bcl-2 was used as a loading control. The bands were assessed and visualized as described in the Materials and Methods. (E) Relative optical density, normalized to Bcl-2, for p-AKT, p-PI3K, p-p38MAPK, Mcl-1 and Bcl-xL immunoblots, n = 5.