Figure 3

Lack of contribution of ATM, ATR and DNA-PK pathways in cisplatin-induced splicing. AT5BIVA (ATM deficient, A) and MO59J (DNA-PK deficient, B) cells were treated with cisplatin (50 μM, 24 hours) and analysed for alternative splicing events in HRNPDL pre-mRNA. (*p ≤ 0.05; **p ≤ 0.01); C-D: MCF7 cells were treated with ATM inhibitor (50 μM) or DNA-PK inhibitor (NU7026; 25 μM) three hours prior to treatment with cisplatin (50 μM for 24 hours); E-F: MCF7 cells were pre-treated with caffeine (5 mM) and with DNA-PK inhibitor (NU7026; 25 μM) for three hours prior to treatment with cisplatin (50 μM) (E: HNRNPDL-E6 p = 0.13; F: AMZ2: p = 0.49). G-L: MCF7 cells were treated with wortmannin (100 nM and 500 nM) or PX866 (500 nM), three hours prior to treatment with cisplatin (50 μM, 24 hours). Modifications of alternative splicing were evaluated for G: HNRNPDL-E6 *p = 0.02; H: HNRNPDL-E8 *p = 0.02; I: AMZ2 **p = 0.004, ***p = 0.0005; J: MDM2. Similar modification was observed with PX866 and illustrated for K: HNRNPDL-E6 *p = 0.05; L: AMZ2 *p = 0.02. Alternative splicing was evaluated by end-point RT-PCR and acrylamide gel electrophoresis. Each bar shows the mean with SD of at least three independent experiments.