Induction of apoptosis by SGI-1776 treatment in MCL cell lines. (A) Dose-dependent induction of SGI-1776–mediated annexin V and PI positivity in MCL cells. MCL cell lines (JeKo-1, Mino, Granta 519, and SP-53) were treated with DMSO alone or with 0.1, 1, 3, 5, or 10μM SGI-1776 for 24 hours, stained with annexin V and PI, and analyzed by flow cytometry. (B) Apoptosis in JeKo-1 and Mino was confirmed by PARP cleavage assay. MCL cells were treated with SGI-1776 for 24 hours in the concentrations mentioned in panel A for immunoblots. (C) Time-dependent induction of SGI-1776–mediated annexin V/PI–positive MCL cells. JeKo-1 and Mino cells were treated with DMSO alone or with 10μM SGI-1776 for 0.5, 1, 2, 4, 8, 16, or 24 hours, stained with annexin V and PI, and analyzed by flow cytometry. (D) Apoptosis in JeKo-1 and Mino was confirmed by PARP cleavage assay. MCL cells were treated with 10μM SGI-1776 for time points mentioned above for immunoblots. All experiments were performed in triplicate.