Alarmin Detection in Senescent Cells

  • Dong Eun Kim
  • Albert R. DavalosEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1896)


Senescent cells secrete diverse array of proteins. One group of proteins, damage-associated molecular pattern (DAMP) proteins exhibit relocalization from inside to outside the cell. High Mobility Group Box 1 protein (HMGB1) is the founding DAMP member. HMGB1 relocalization from the nucleus provides a molecular signature during senescence. We provide distinct molecular techniques (immunofluorescence, immunohistochemistry, and Western blot assays) to assess HMGB1 relocalization during the initial stages of senescence.

Key words

Damage-associated molecular pattern (DAMP) Alarmin High Mobility Group Box 1 protein (HMGB1) Immunofluorescence Immunohistochemistry Protein precipitation Western blot 


  1. 1.
    Bianchi ME (2007) DAMPs, PAMPs and Alarmins: all we need to know about danger. J Leukoc Biol 81:1–5CrossRefGoogle Scholar
  2. 2.
    Rider P, Voronov E, Dinarello CA, Apte RN, Cohen I (2017) Alarmins: feel the stress. J Immunol 198(4):1395–1402CrossRefGoogle Scholar
  3. 3.
    Coppé JP, Patil CK, Rodier F, Sun Y, Muñoz DP, Goldstein J, Nelson PS, Desprez PY, Campisi J (2008) Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol 6:2853–2868CrossRefGoogle Scholar
  4. 4.
    Kuilman T, Michaloglou C, Vredeveld LCW, Douma S, van Doorn R, Desmet CJ, Aarden LA, Mooi WJ, Peeper DS (2008) Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell 133(6):1019–1031CrossRefGoogle Scholar
  5. 5.
    Davalos AR, Kawahara M, Malhotra GK, Schaum N, Huang J, Ved U, Beausejour CM, Coppe JP, Rodier F, Campisi J (2013) p53-dependent release of Alarmin HMGB1 is a central mediator of senescent phenotypes. J Cell Biol 201:613–629CrossRefGoogle Scholar
  6. 6.
    Jeon OK, Kim C, Laberge RM, Demaria M, Rathod R, Vasserot AP, Chung JW, Kim DH, Poon D, David N, Baker DJ, van Deursen JM, Campisi J, Elisseeff JH (2017) Local clearance of senescent cells attenuates the development of post-traumatic osteoarthritis and creates a pro-regenerative environment. Nature Med 23(6):775–781CrossRefGoogle Scholar
  7. 7.
    Wiley CD, Velarde MC, Lecot P, Liu S, Sarnoski EA, Freund A, Shirakawa K, Lim HW, Davis SS, Ramanathan A, Gerencser AA, Verdin E, Campisi J (2016) Mitochondrial dysfunction induces senescence with a distinct secretory phenotype. Cell Metab 23(2):303–314CrossRefGoogle Scholar
  8. 8.
    Pandolfi F, Altamura S, Frosali S, Conti P (2016) Key role of DAMP in inflammation, cancer, and tissue repair. Clin Ther 38(5):1017–1028CrossRefGoogle Scholar
  9. 9.
    Vénéreau E, Ceriotti C, Bianchi M (2015) DAMPs from cell death to new life. Front Immunol 18(6):422Google Scholar
  10. 10.
    He SJ, Cheng J, Feng X, Yu Y, Tian L, Huang Q (2017) The dual role and therapeutic potential of high-mobility group box 1 in cancer. Oncotarget 8(38):64534–64550PubMedPubMedCentralGoogle Scholar
  11. 11.
    Tang D, Shi Y, Kang R, Li T, Xiao W, Wang W, Xiao X (2006) Hydrogen peroxide stimulates macrophages and monocytes to actively release HMGB1. J Leukoc Biol 81(3):741–747CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Buck Institute for Research on AgingNovatoUSA

Personalised recommendations