Figure 4

Down-regulation of survivin changes the dynamics of microtubule networks. (A) Supernatant and pellet fractions of un-treated KB and KB-L30 cells were extracted. Proteins were resolved by SDS-PAGE and analyzed by western blotting. Anti-actin was used as an internal control. Relative band intensities were shown. (B and C) KB and KB-L30 cells were transfected with siR-C (scramble control) and siR-S (survivin specific) for 48 h. Total lysate and pellet fractions of cells were extracted. Proteins were resolved by SDS-PAGE and analyzed by western blotting. Anti-actin was used as an internal control. Relative band intensity was shown. (D) Down-regulation of survivin reduced tubulin acetylation in KB and KB-L30 cells. Cells were transfected with siR-C or siR-S for 24 h and co-incubated with/without 8 nM (KB) and 70 nM (KB-L30) of BPR0L075 for 48 h. Total cell lysate of cells was extracted. Proteins were resolved by SDS-PAGE and analyzed by western blotting. Anti-actin was used as an internal control. (E) Down-regulation of survivin destabilizes the microtubule networks. KB cells were seeded on 8-well chamber slides. Cells were pre-transfected with siR-C (scramble control) or siR-S (survivin specific) for 24 h and co-incubated with 4 nM of BPR0L075 for 24 h. Cells were fixed and labeled with mouse anti-human α-tubulin antibody and FITC-coupled anti-mouse secondary antibody. Slides were analyzed by fluorescent microscopy.