Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting of tubulin polymerization and induction of mitotic blockage by Methyl 2-(5-fluoro-2-hydroxyphenyl)-1 H -benzo[ d ]imidazole-5-carboxylate (MBIC) in human cervical cancer HeLa cell

doi: 10.1186/s13046-016-0332-0

Figure Lengend Snippet: a MBIC disrupts mitotic spindle: HeLa cells expressing EGFP-α-tubulin, EGFP-CENP-A and histone H2B-mCherry were treated with DMSO (upper), MBIC (10 μM, middle), or nocodazole (2 μM, lower) and imaged at 15 min intervals. Time (min) is indicated in the upper left of each panel. For middle and lower panels, arrowheads mark the cell in the center at the first frame. Scale bar = 5 μm. b Effect of MBIC on tubulin polymerization: Tubulin polymerization assay was conducted using Tubulin polymerization assay kit (Cytoskeleton, Inc.). The plate was read using Infinite® 200 PRO—Tecan regulated on 96 well plate reader spectrophotometer. The plate was read at 340 nm in kinetic mode for two hours. Figure 3 shows curves of tubulin treated by paclitaxel (10 μM), nocodazole (10 μM), colchicine (10 μM), MBIC (10 μM) and untreated tubulin. Maximal velocity (Vmax) of each drug on tubulin polymerization was calculated

Article Snippet: Samples were mixed well and the tubulin assembly was monitored by an increase in fluorescence emission at 340 nm in kinetic mode for 120 min at 37 °C using a plate reader (Infinite® 200 PRO—Tecan plate reader, USA).

Techniques: Expressing, Polymerization Assay, Spectrophotometry