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ATM: - WT KD

Supplementary Figure S1. Sequences of amino acids surrounding potential ATM phosphorylation sites on B56γ isoforms and B56δ. ATM: - WT KD. B56  Phos. HA-B56 . FLAG-ATM. p53 S15 Phos. p53. vinc. 0 1h 2h 3h 4h 6h 8h 12h. -. B56 γ 3. IR. -. S510A. IR. -.

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ATM: - WT KD

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  1. Supplementary Figure S1. Sequences of amino acids surrounding potential ATM phosphorylation sites on B56γ isoforms and B56δ.

  2. ATM: - WT KD B56 Phos HA-B56 FLAG-ATM p53 S15 Phos p53 vinc

  3. 0 1h 2h 3h 4h 6h 8h 12h - B56γ3 IR - S510A IR - S510D IR Supplementary Figure S3. Loading controls for Figure 2A. Whole cell lysates from U2OS cells transfected with the HA-tagged B56γ construct listed, treated with cyclohexamide, then either mock treated (-) or IR treated, and harvested at various time points, were subjected to western blot with anti-vinculin antibody.

  4. Cyclohexamide: 0h 1h 2h 4h 6h 8h B56γ3 B56γ3 vinc B56γ3 B56γ3 + MDM2 vinc B56γ3 B56γ3 + C464A vinc S510A S510A vinc S510A S510A + MDM2 vinc S510D S510D vinc S510D S510D + MDM2 vinc Supplementary Figure S4. Ser510 phosphorylation blocks the ability of MDM2 expression to shorten B563 half life. U2OS cells were tranfected with either S510A, S510D, or wild type B563, and either wild type MDM2, C464A mutant, or an empty vector control, then treated with cyclohexamide, harvested at the time points shown, and analyzed for B563 protein levels by HA immunoblotting. Vinc: vinculin.

  5. Input B56γ-IP IgG CycG RNAi: - + - + - + MDM2 B56γ p53 PP2A A Cyclin G vinc Supplementary Figure S5. Cyclin G knock down has no effect on B56 interaction with MDM2. U2OS cells were subjected to RNAi knock down of Cyclin G (+) or control treatment (-), then lysed and analyzed for interaction with B56 by immunoprecipitation followed by blotting with the antibodies listed. Vinc: vinculin.

  6. Cont CycG RNAi MDM2: - WT CA - WT CA HA-B56γ3 MDM2 Cyclin G vinc Supplementary Figure S6. MDM2-mediated decrease in B563 protein levels is not affected by Cyclin G. U2OS cells were subjected to RNAi knock down of Cyclin G (CycG RNAi), or control treatment (Cont), along with transfection of B563 protein, and either wild type or C464A mutant MDM2 (CA), or an empty vector control (-), and subjected to western blot to detect levels of B563 expression, as well as the other proteins listed. Vinc: vinculin.

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