A Phthalimide Derivative That Inhibits Centrosomal Clustering Is Effective on Multiple Myeloma
Figure 7
NPM knockdown induces multipolar spindles.
HeLa cells were transfected with siRNA for luciferase (control) or NPM. (A) After 48 h, the whole cell lysates were analyzed by western blotting with antibody against NPM (left) and the band intensities were quantified (right). (B) Immunofluorescence with anti-NPM (green) and anti-γ-tubulin (red) antibody was performed (left). White arrows indicate centrosomes. At least 50 mitotic cells were counted in three independent experiments. The ratio of cells with multipolar spindles under the indicated conditions was quantified (below). (C) Representative multinucleated cell under the indicated conditions. (D) After 72 h, caspase-9 activity was determined using luminescence-based assay. (E) At 48 h after siRNA transfection, the cells were treated with 0–2.5 µM TC11 and then incubated for a further 72 h. Cell viability was determined by means of WST-1 assay in three independent experiments. An asterisk denotes a statistically significant difference according to Student’s t-test (P<0.05). Bar; 10 µm.