Lyn aberrant activity generates varying tyrosine phosphorylation patterns of the cytosolic subproteome in B-CLL cells. (A) Cytosol of freshly isolated B cells from normal donors and B-CLL patients distributed between the UM-CLL (patients 4, 15, and 34) and M-CLL (patients 11, 25, and 38) subsets was analyzed by Wb analysis with anti-pTyr antibody and probed with anti-β-actin antibody as a loading control. Asterisks indicate recurrent bands within the B-CLL samples. (B) Cytosol of freshly isolated B-CLL cells, purified from patients in different clinical stages and belonging to the unmutated (UM-CLL) and mutated (M-CLL) IgVH subsets (patients 4 and 25, respectively, in this figure) and cultured in the absence (left panels) or presence (right panels) of 10 μM PP2, was separated by 2D-PAGE using a linear wide range (pH 3-10) immobilized pH gradient in the first dimension for subsequent Wb analysis with anti-pTyr antibody. The figure is representative of samples from 6 normal donors and 24 CLL patients belonging to UM-CLL and M-CLL subsets, each of which comprised 12 samples. IEF, isoelectric focusing; SDS/PAGE, sodium dodecyl sulfate/polyacrylamide gel electrophoresis.
