Maltonis induces DNA damage and triggers apoptosis in sarcoma cell lines. (A) Induction of H2AX phosphorylation (γ-H2AX) in TC-71 cells after exposure for 24 hours to 3 μM maltonis (magnification × 40) (B) Evaluation of nuclear fragmentation after staining with HOECHST-33258 on untreated or 3, 10 μM treated RD/18, U-2OS or TC-71 cells after 72 h. Histogram shows mean percentages ± SE of fragmented nuclei out of 1000 considered in three independent experiments. Statistical analysis was performed by Student’s t test: *P < 0.05. Representative pictures are also shown (magnification × 400). (C) Apoptosis-related DNA fragmentation induced by maltonis exposure after 72 hours at 10 or 25 μM in TC-71 cells. (D) Western blotting on TC-71 total cell lysates from control or maltonis treated cells (3 μM for 72 h) showing caspase 3 and PARP cleavage as well as BAX induction. Equal loading was monitored with anti-GAPDH or beta-actin blotting (lower lines).