SDF-1 mRNA expression by NLC and p44/42 mitogen-activated protein kinase (Erk 1/2) activation in CLL B cells by SDF-1α.

(A) cDNA from purified NLC from 4 patients with CLL was examined for the expression of SDF-1 mRNA by RT-PCR. The specific 230-bp PCR fragment is visible in each of the 4 NLC samples (lanes 2-5), and in the positive control, namely an SDF-1–containing plasmid (lane 6). Lanes 7-11 display PCR fragments of the expected size using GA3PD primers for the NLC samples (lanes 7-10) but not for the SDF-1 plasmid control. Lanes 1 and 12 display the separation of the 100-bp marker DNA, and the 200, 600, and 1000 bp bands are marked on the left side. (B) p44/42 MAPK activation in CLL B cells treated with SDF-1α at 200 ng/mL. CLL cell lysates were obtained at the time points indicated on the horizontal axis and examined for phospho-p44/42 MAPK protein by Western blot analysis. Protein bands of the expected sizes of 42 and 44 kd, as indicated on the left, were prominent after the stimulation of CLL cells with SDF-1α, whereas they were only weakly apparent before activation by SDF-1 (time 0).