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. 2009 Nov 24;38(3):832–845. doi: 10.1093/nar/gkp1039

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© The Author(s) 2009. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — APE1 and SIRT1 associate with each other. (A) APE1 and SIRT1 bind to each other in HEK 293 cells. Immunoprecipitation of epitope-tagged SIRT1 co-precipitates epitope-tagged APE1 expressed in HEK 293 cells. (B) APE1 does not bind to catalytically inactive dominant negative SIRT1. Immunoprecipitation of epitope-tagged APE1 co-precipitates epitope-tagged wild-type SIRT1 but not dominant negative SIRT1 (H363Y) expressed in HEK 293 cells. (C) Endogenous SIRT1 binds to endogenous APE1. Co-immunoprecipitation of endogenous SIRT1 and APE1 in HEK 293 cells. (D) Co-localization of fluorescent epitope-tagged SIRT1 and APE1 expressed in HEK 293 cells. Co-localization of extra-nucleolar (white arrow) APE1 but not nucleolar (blue arrow) APE1 with SIRT1 is shown. (E) Hydrogen peroxide (H₂O₂) promotes binding of APE1 to SIRT1. H₂O₂ (500 µM, 30 min) increases co-precipitation of epitope-tagged SIRT1 in immunoprecipitates of epitope-tagged APE1 expressed in HEK 293 cells. WCL: whole cell lysate. N-IgG: non-immune immunoglobulin. FLAG-APE1 and Myc-SIRT1 were expressed in A and B, DsRed-APE1 and EGFP-SIRT1 in C, and DsRed-APE1 and Myc-SIRT1 in D.