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. 2016 Oct 7;14(6):804–813. doi: 10.1080/15476286.2016.1243647

Figure 2.

Figure 2.

ATM phosphorylates WRAP53β upon IR and UV exposure. (A) U2OS cells stably overexpressing Flag-WRAP53β were irradiated (6 Gy) and harvested at the time-points indicated for western blotting with the antibodies indicated. (B) U2OS cells stably expressing Flag-WRAP53β were treated with the inhibitors indicated for 16 h or siWRAP53 for 48 h, exposed to 6 Gy IR, and harvested 1 h later for western blotting with the antibodies indicated. (C) U2OS cells stably expressing Flag-WRAP53β were treated with the inhibitors indicated for 24 h or siWRAP53 for 48 h, exposed to 30 J/m2 UV, and harvested 3 h later for western blotting with the antibodies indicated. (D) U2OS cells stably overexpressing Flag-WRAP53β were exposed to 30 J/m2 UV and harvested at the time-points indicated for western blotting with the indicated antibodies. (E) U2OS cells stably expressing Flag-WRAP53β were treated with the siRNAs indicated for 48 h or an inhibitor of ATM (ATMi) for 24 h, exposed to 6 Gy IR, and harvested 1 h later for western blotting with the antibodies indicated. (F) Parental U2OS cells were exposed to 6 Gy IR (1 hour recovery) or 30 J/m2 UV (9 hours recovery) and harvested for western blotting with the antibodies indicated. H1299 cells were treated with ATMi for 16 h or siWRAP53 for 48 h, irradiated with 6 Gy and harvested 30 min later for western blotting of pWRAP53βS64, WRAP53β and β-actin.