Supplementary Materials

Supplementary Materials for:

Inflammatory stimuli induce inhibitory S-nitrosylation of the deacetylase SIRT1 to increase acetylation and activation of p53 and p65

Shohei Shinozaki, Kyungho Chang, Michihiro Sakai, Nobuyuki Shimizu, Marina Yamada, Tomokazu Tanaka, Harumasa Nakazawa, Fumito Ichinose, Yoshitsugu Yamada, Akihito Ishigami, Hideki Ito, Yasuyoshi Ouchi, Marlene E. Starr, Hiroshi Saito, Kentaro Shimokado, Jonathan S. Stamler, Masao Kaneki*

*Corresponding author. E-mail: mkaneki{at}helix.mgh.harvard.edu

This PDF file includes:

  • Fig. S1. Mutation of the CXXC motif in SIRT1 abolishes its ADP-ribosyltransferase activity, and SNAP decreases SIRT1 deacetylase activity.
  • Fig. S2. S-Nitrosylation is a major thiol modification of SIRT1 in cultured cells exposed to SNAP and carmustine.
  • Fig. S3. The specific detection of SIRT1 S-nitrosylation is confirmed by the effect of preincubation with HgCl2 and by ascorbate-dependent biotinylation.
  • Fig. S4. TPEN blocks reactivation of NO donor–exposed recombinant SIRT1 by DTT.
  • Fig. S5. SIRT1M3 decreases cell viability.
  • Fig. S6. NO donors increase the abundance of acetylated p53.
  • Fig. S7. GSNO and SNAP decrease viable cell numbers in a p53-dependent manner.
  • Fig. S8. GSNO induces p53 acetylation and decreases viable cell numbers independent of cGMP.
  • Fig. S9. Inhibition of iNOS prevents the ability of inflammatory molecules or iNOS overexpression to induce S-nitrosylation of SIRT1 and acetylation of p53.
  • Fig. S10. Inhibition of iNOS prevents the ability of inflammatory molecules to induce acetylation of p65.
  • Fig. S11. iNOS deficiency prevents the ability of LPS to induce acetylation and activation of p53 and p65 in mouse liver.
  • Fig. S12. Inhibition of nNOS prevents the ability of MPTP to induce acetylation and activation of p53 and p65 in the nigra-striatum of MPTP-injected mice.
  • Fig. S13. Acetylation and activation of p53 and p65 are increased in skeletal muscle of aged rats compared to young rats.
  • Fig. S14. Inhibition of iNOS decreases acetylation and activation of p53 and p65 in skeletal muscle of aged rats.

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Citation: S. Shinozaki, K. Chang, M. Sakai, N. Shimizu, M. Yamada, T. Tanaka, H. Nakazawa, F. Ichinose, Y. Yamada, A. Ishigami, H. Ito, Y. Ouchi, M. E. Starr, H. Saito, K. Shimokado, J. S. Stamler, M. Kaneki, Inflammatory stimuli induce inhibitory S-nitrosylation of the deacetylase SIRT1 to increase acetylation and activation of p53 and p65. Sci. Signal. 7, ra106 (2014).

© 2014 American Association for the Advancement of Science