Figure 2

Down-regulation of survivin in KB and KB- L30 cells by siRNA. (A) Cells were seeded on 24-well plates overnight and transfected with siR-C (scramble control) or siR-S (survivin specific) for 36 h. Total RNA was extracted by Trizol reagent. RT-PCR was performed and PCR products were resolved by DNA electrophoresis. GAPDH was used as an internal control (B) Cells were seeded on 8-well chamber slides overnight and transfected with siR-C (scramble control) or siR-S (survivin specific) for 48 h. Cells were labeled with FITC-coupled anti-human survivin antibody and counter-stained with DAPI nucleus stain. Slides were analyzed using fluorescent microscopy. (C) Cells were seeded on 6-well plates overnight and transfected with siR-C or siR-S for 48 h. Cell lysate were extracted and proteins were resolved by SDS-PAGE. Western blot analysis was performed as described in Materials and Methods.