siRNA knockdown of TRIP-Br2 expression inhibits cell-autonomous growth of HCT-116 cells. 4 pmol or 40 pmol of Cy3-labeled oligomer control (Cy3-O), scrambled siRNA (Scr) and TRIP-Br2- specific siRNAs (DS1, DS2 or DS3) were transiently transfected into HCT-116 cells respectively. "Not transfected" (NT) samples were used as negative controls. The specificity of TRIP-Br2-specific siRNAs was assessed by semi-quantitative RT-PCR and Western blot analyses. Three independent experiments were performed in triplicates. (A) Knockdown of TRIP-Br2 expression in HCT-116 cells (12-well plate) was achieved by TRIP-Br2- specific siRNAs, DS1 and DS2, at the dose of 40 pmol. 18srRNA was used as a loading control. (B) TRIP-Br2 protein expression was significantly knocked down by TRIP-Br2-specific siRNAs, DS1 and DS2, at the dose of 40 pmol. β-tubulin was used as a loading control. (C) Colony forming assays and (D) cell count analyses showed that siRNA knockdown of TRIP-Br2 expression (DS-1 or DS-2 at the dose of 40 pmol) inhibited cell-autonomous growth of serum-deprived HCT-116 cells. The dashed line indicates the initial cell density plated. Data was obtained from three independent experiments that were performed in triplicates.