Fig. 1: ITF2357 affects survival of human sarcoma cells.

a Analysis of cell viability by MTT assay in the indicated sarcoma cell lines treated for 72 h with increasing concentrations of ITF2357. The results are reported as “viability of drug-treated cells/viability of control cells” × 100 and represent the mean ± SD of three independent experiments performed in triplicate. b Colony formation of HT1080 cells treated for 24 h with increasing concentrations of ITF2357. Representative images are shown. c Western blot analysis of acetylated histone H3 (Ac-H3), acetylated α-Tubulin (Ac-Tubulin) in total cell lysates from the indicated cell lines treated with 0.1 and 0.5 μM ITF2357 for 24 h. d Representative experiment of apoptotic cells evaluation by AnnexinV/PI staining in the indicated cell lines exposed to increasing concentration of ITF2357 for 72 h. e Quantification of apoptosis by AnnexinV/PI staining in the indicated cell lines treated for 72 h with increasing concentrations of ITF2357. The results represent the mean ± SD of three independent experiments. f Western blot analysis of cleaved form of PARP (PARP cl.) in total cell lysates from HT1080 cells treated with 1 and 5 μM ITF2357 for 72 h. c, f HSP72/73 expression was used as loading and transferring control. Western blots representative of two independent experiments with similar results are shown. g Flow cytometric analysis of active caspase-3-PE staining in HT1080 cells untreated or treated for 72 h with 1 μM ITF2357 alone or in combination with 50 μM pan-caspase inhibitor zVAD-fmk (zVAD). A representative experiment out of two is shown. a, e p values were calculated between untreated and treated cells, *p < 0.01