Figure 5
Effect of KUD773 on microtubule assembly assay. (A) PC-3 cells were incubated in the absence or presence of the indicated agent (taxol, 0.1 μM; vincristine, 0.1 μM). The cells were harvested and separated into soluble (S, microtubule disassembly) and particulate form (P, microtubule assembly), and proteins were detected by Western blot analysis. The data for relative expression of particulate fraction are also demonstrated. (B) Cells were incubated with or without the indicated agent for 12 h. The cells were fixed for the confocal microscopic detection of microtubule organization by staining with anti-β-tubulin antibody (green fluorescence) and DAPI (blue fluorescence, for nuclear detection). (C) Cells were treated in the same condition to (B) but the detection of microtubule organization by staining with anti-γ-tubulin antibody (green fluorescence) and DAPI (blue fluorescence). Arrowhead, monopolar γ-tubulin staining. Scale bar, 20 μm. (D) Aurora A luminescent kinase assay was performed. The data are expressed as mean ± SEM of three independent experiments.