Reduction of global RNA synthesis and Mcl-1 mRNA and protein expression in MCL cell lines by SGI-1776. (A) Dose-dependent effects of SGI-1776 treatment on global transcription level in JeKo-1, Mino, Granta 519, and SP-53 cells. Cells were treated with DMSO alone or with 0.1, 1, 3, 5, or 10μM SGI-1776 for 24 hours and then incubated with uridine for 30 minutes, and radioactive incorporation was measured via scintillation counter. (B) Time-dependent effects of SGI-1776 treatment on global transcription level in JeKo-1 and Mino cells. Cells were treated with DMSO alone or with 10μM SGI-1776 for 0.5, 1, 2, 4, 8, 16, or 24 hours and then prepared for uridine incorporation as described in panel A. Dose-dependent (C) and time-dependent (D) effects of SGI-1776 on MCL1 mRNA expressions in JeKo-1 and Mino cells. Cells were treated with SGI-1776 in dose-dependent or time-dependent manner as mentioned in Panels A and B, and RNA was isolated and analyzed using real-time RT-PCR. Impact of SGI-1776 in dose-dependent (E) and time-dependent (F) treatments on Mcl-1 protein expression in JeKo-1 and Mino cells. Mcl-1 protein expression was measured by immunoblots in both dose- and time-dependent SGI-1776 treatments (see panels A and B) of JeKo-1 and Mino cells. Quantitation of immunoblot with dose-dependent SGI-1776 treatment in JeKo-1 (G) and Mino (H) cells. Mcl-1-to-GAPDH ratio was calculated, and data are presented as means of 3 independent experiments ± SEM.