Ku 55 933 s1092

Specific antibodies against p53 (DO-1, sc-126), p15 (D-12, sc-271,791), p19 (SPM429, sc-65,594) and RanBPM (sc-271,727) antibodies were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA), and p18 (#2896), p21 (#2947), p57 (#2557) and RanBPM (#14,638) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-p16 (A301-267 A) was purchased from Bethyl Laboratories (Montgomery, TX, USA). Anti-p27 (AF1669), anti-ATM (AF1399) and anti-phospho-ATM (Ser1981) were purchased from Beyotime Biotechnology (Shanghai, China). Anti-USP11 (ab109232) and anti-RanBPM (ab205954) antibodies were purchased from Abcam (Shanghai, China). Anti-GAPDH (AT0002) antibodies were purchased from CMCTAG (Milwaukee, WI, USA). Anti-Flag (cat. M185-3 L), anti-Myc (cat. M192-3) and anti-HA (cat. M180-3) antibodies were purchased from Medical & Biological Laboratories CO., LTD. (Minato-ku, Tokyo, Japan). Etoposide (VP-16, S1225) and KU-55,933 (S1092) were purchased from Selleck Chemicals (Shanghai, China), and Cycloheximide (sc-3508B) and MG132 (C2211) were purchased from Sigma (Shanghai, China). Doxorubicin (SC0159) was purchased from Beyotime Biotechnology (Shanghai, China).

The human ovarian cancer cell lines, SK-OV-3 and A2780S, and PAC-resistant A2780T were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in DMEM, supplemented with 10% fetal bovine serum and 50 U/ml penicillin and streptomycin.
MTT, DMSO, Hoechst 33258, PAC and anti-β-actin, Bad and Bcl-2 were purchased from Sigma (St Louis, MO, USA). JSH-23 (S7351), SC75741 (S7273), CAPE (S7414) and KU55933 (S1092) were bought from Selleckchem (USA). Antibodies against Bax, ATM, p-ATM, IKKα, p-IKKα/β, IκBα, p-IκBα, NF-κB (p65), H2AX and γ-H2AX were obtained from Cell Signaling Technology (CST, Danvers, MA). Antibody against Bcl-xl was bought form Abcam (UK). MIL was isolated from Millettia pachycarpa Benth. The Sample in all experiments contained MIL of 99% or higher purity, which was dissolved in DMSO to produce a 10 mM stock solution and stored at 4°C. In the in vivo experiments, MIL was dissolved in sodium chloride injection containing 2.5% Tween-80 and 2.5% ethanol in a volume of 200 μL.