Confirmation of the siRNA screening results. (A) Validation of the effects of STK10, TNK2 and PLK1 silencing on cell growth of TC-32, TC-71, SK-ES1 and RD-ES cells using two siRNAs per gene. Ewing's sarcoma cells were transfected with siRNA by reverse transfection in 384-well microtiter plates and grown for 96 hrs. Each siRNA treatment was performed in quadruplicate wells. Cell number was assessed as described in Material and Methods. Treatment of Ewing's sarcoma cells with siRNA targeting STK10, TNK2 and PLK1 significantly decreased cell viability (p < 0.0001) compared to non-silencing siRNA controls. Data is representative of three independent experiments. (B) Representative images of TC-32 and TC-71 Ewing's sarcoma cells treated with siRNA shows decreased cell number and increased cellular toxicity. Bar represents 100 μm.