Fig. 6.

Biological effects of treatment with EIF4A3 inhibitor, T-595. a Treatment with T-595 at high doses or with siRNA to EIF4A3 and after 48 h results in G2M arrest and sub-G1/G0 fraction indicative of apoptosis. Cell cycle profiles (counts (vertical axes) vs DNA content (horizontal axes)) were assessed by flow cytometry after the treatment DMSO, 1 or 5 μM T-595 or siRNA (siNT#1, siEIF4A3#2,#3) for the respective times shown. The percentage of cells assigned to be in the respective stages of the cell cycle is indicated on the respective charts. b Apoptosis induced on treatment with eutomer, T-595 but not distomer, T-598. Graphs showing apoptosis as assessed by Caspase 3/7 activity (vertical axes) when HeLa cells were treated for 24 h with various concentrations of T-595 (blue) or T-598 (red) (horizontal axes). c Microscopic visualization of spindle and chromosome proteins reveals chromosome mis-segregation. Representative images of untreated (top panel) and treated (5 μM T-595 for 30 h, bottom two panels) showing chromosomal DNA stained with DAPI (blue), centromere proteins stained with anti-CENP-B (red) or microtubules identified by anti-α-tubulin (green). Scale bar = 10 μm