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. Author manuscript; available in PMC: 2011 Oct 25.
Published in final edited form as: Nat Cell Biol. 2007 Oct 14;9(11):1253–1262. doi: 10.1038/ncb1645

Figure 1.

Figure 1

SIRT1 is sumoylated in vivo and in vitro. (a) DU145 cell extracts were immunoprecipitated (IP) with antibodies against Flag or SIRT1 and immunoblotted (IB) with antibodies against Sumo-1 or SIRT1, as indicated. S-SIRT1, sumoylated SIRT1. (b) Cell extracts were immunoprecipitated with antibodies against Flag or Sumo-1 and immunoblotted with antibodies against SIRT1 or Sumo-1, as indicated. Note that the antibody against SIRT1 detected three proteins in the input lane. This antibody is known to detect a non-specific protein with an Mr of approximately 100K in western blots. (c) Cells expressing His–Sumo-1 were extracted under denaturing conditions. His–Sumo-1 conjugates were isolated with Ni-NTA agarose and immunoblotted with antibodies against SIRT1. The error bars represent s.d. (d) Sumoylation reactions using recombinant human SIRT1 from bacteria were carried out in the presence of the indicated components. Reaction mixtures were immunoblotted with antibodies against SIRT1. Uncropped images of the scans are shown in the Supplementary Information, Fig. S1.