Effect of Nck2 on human metastatic melanoma cell proliferation. (A) Total cell lysates (30 μg protein) prepared from 451Lu human melanoma cells transfected with control or specific Nck2 siRNA were subjected to western blot analysis using specific antibodies that recognize Nck1 or Nck2 (left panel). RT-PCR was performed to determine Nck1 and Nck2 mRNA levels using total RNA and specific primers for human Nck1 and Nck2. 18S was used as control (right panel). (B) 451Lu human melanoma cells transfected with control or Nck2 siRNA were followed for proliferation 1 and 2 days after siRNA transfection. Results represent cell density corresponding to the mean of Crystal Violet incorporation ± SD from one experiment performed in triplicate. * p < 0.05 vs control siRNA using Student's t-test. Similar results were found in three independent experiments.