Fig. 5.

RANBP9 ablation impairs ATM activation and results in increased amounts of chromatin-bound PARylated proteins. a WB analysis of RANBP9 WT and KO clones left untreated or after 24 h of treatment with 20 μM CDDP, using the indicated antibodies. GAPDH was used as loading control. b Cell cycle analysis by flow cytometry of propidium iodide stained RANBP9 WT and KO cells, not treated or treated with 30 μM CDDP for 48 h. Red peaks indicate G1 and G2 phase, stripes indicate S phase. After the treatment with the platinum compound, RANBP9 WT cells exhibit G2 phase blockage, while RANBP9 KO clones are blocked in S phase. c WB analysis of RANBP9 WT and KO clones left untreated or after 24 h of treatment with 20 μM CDDP or 30 min of 100 nM insulin using the indicated antibodies. GAPDH was used as loading control. d PARylation levels in RANBP9 WT and KO clones after the treatment with CDDP evaluated by Western blot. Cells were left untreated or exposed to 20 μM CDDP for 30 min. Then chromatin bound and nuclear soluble fractionated proteins were extracted. Histone H3 was used as loading control for chromatin bound fraction, nucleophosmin (B23) for nuclear soluble proteins