Fig. 2.

On-target validation of PAK5 in H2087 cells. (A and B) PAK5 was knocked down in H2087 (SMARTpool 100 nM; oligos 1–4, 50 nM). Plates were subjected to (A) BrdU assay and (B) caspase 3/7 activity apoptosis assay. Data (n = 9) were normalized to nontargeting siRNA (NEG). Error bars indicate SD. P values were calculated by using one-tailed paired Student t test (*P < 0.005, **P < 0.01, and ***P < 0.05 vs. nontargeting siRNA control). (C) On-target effect of PAK5 SMARTpool (100 nM) and individual oligos (50 nM) was also verified by Western blotting. (D) H2087 cells were reverse-transfected with 100 nM siRNA as indicated (BRAF and SMARTpool siNRAS and siPAK5). Knockdown of PAK5 was confirmed by RT-PCR.