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. Author manuscript; available in PMC: 2016 Mar 9.
Published in final edited form as: Cell Rep. 2016 Feb 4;14(6):1435–1447. doi: 10.1016/j.celrep.2016.01.018

Figure 3. SIRT2 Deacetylation of ATRIP at K32 Promotes ATR Activation.

Figure 3

(A) HCT116 cells were transfected with NT or SIRT2 siRNA, treated with or without HU, harvested, immunoprecipitated with an anti-ATR antibody, separated by SDS-PAGE, and immunoblotted with antibodies against P-ATR T1989, ATR, SIRT2 and GAPDH. (B) HCT116 cells were transfected with NT or SIRT2 siRNA, treated with or without HU, harvested, separated by SDS-PAGE, and immunoblotted with indicated antibodies. (C) Cells stably expressing an empty vector or FLAG-SIRT2 and transfected with NT or SIRT2 siRNA targeting the 3′UTR, were treated with or without HU, harvested, separated by SDS-PAGE, and immunoblotted with indicated antibodies. (D) 293T cells were transfected with ATRIP WT or K32Q, K96Q, or K101Q, treated with or without HU, immunoprecipitated with anti-HA beads, separated by SDS-PAGE, and immunoblotted with antibodies against HA, P-ATR T1989 and ATR. (E) 293T cells were transfected with ATRIP WT or K32R, or K32Q, treated with or without HU, and extracted for nuclear extraction. The nuclear extraction was separated by SDS-PAGE, and immunoblotted with antibodies against HA, P-ATR T1989 and ATR. (F) HCT-116 cells stably expressing an empty vector or HA-ATRIP WT or K32Q and transfected with NT or SIRT2 siRNA, were treated with or without HU, harvested, separated by SDS-PAGE, and immunoblotted with indicated antibodies. See also Figure S3.