Inhibition of SHP-2 expression by SHP-2 antisense oligonucleotides or siRNA decreases p210 level. (A) p210-expressing K562 leukemia cells were transfected with SHP-2 antisense, sense, and nonsense oligonucleotides (7.5 μM) using the Fugene transfection reagent. Cells were harvested 8 hours after transfection and examined for p210 with anti–c-Abl IB. The blot was stripped and reimmunoblotted with anti–SHP-2 and anti-Erk Abs. (B) Cell-survival rates were determined by trypan blue exclusion assay. The asterisk indicates significant differences between antisense versus sense or nonsense (P < .01 by Student t test). (C) WT/Bcr-Abl cells were transfected with SHP-2 siRNA by electroporation (25 μg for 1 × 107 cells). Cells were harvested 12 hours after transfection and analyzed for p210 level with anti–c-Abl IB. The blot was stripped and reimmunoblotted with anti–SHP-2 and anti-Erk Abs. (D). Fold changes in cell numbers of siRNA-transfected cells over 3 days of culture were determined by the MTS assay as described in Figure 1A. Results displayed are representative of 2 to 3 independent experiments. Results shown on panels B and D are the mean ± SD of triplicates from one experiment.
