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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Mol Cell. 2021 Feb 10;81(7):1515–1533.e5. doi: 10.1016/j.molcel.2021.01.019

Figure 1. PARylation promotes protein aggregation in human cells lacking ATM.

Figure 1.

(A) Human U2OS cells were grown with ATM shRNA-mediated depletion or mock treatment (3 biological replicates for each), all with arsenite (25 μM). Cell lysates were analyzed by mass spectrometry, identifying 2363 polypeptides present in all samples. Levels of each protein in control versus ATM-depleted cells are shown, with non-significant differences in grey and significant differences in red, after FDR control at 0.05. (B) Detergent-resistant aggregates were prepared from control and ATM-depleted cells as described in (A). (C) U2OS cells with control or ATM shRNA were treated with veliparib (10 μM) and arsenite (25 μM) as indicated. Lysates and detergent-resistant aggregates were analyzed by western blotting for CK2β or PSMB2. (D, E) Levels of CK2β (D) or PSMB2 (E) in aggregate fractions normalized by lysate levels were quantified in three replicates of (C); shown relative to control cells. (F) ATM-depleted U2OS cells were grown with doxycycline-induced PARP1 shRNA-mediated depletion or mock treatment as indicated, all with arsenite (25 μM). Levels of ATM, PARP1, CK2β and PSMB2 in lysates and aggregate fractions were analyzed by western blotting. (G, H) Levels of CK2β (G) or PSMB2 (H) in aggregate fractions normalized by lysate levels were quantified in three replicates of (F); shown relative to control cells. *, **, ***, and **** indicate p<0.05, 0.005, and 0.0005 by Student two-tailed t-test; NS = not significant.