Research Paper Volume 11, Issue 2 pp 549—572

S100A13 promotes senescence-associated secretory phenotype and cellular senescence via modulation of non-classical secretion of IL-1α

Figure 3. Inhibition the binding of IL-1α to S100A13 or to Cu2+ suppresses IL-1α secretion, NF-κB activity, and SASP expression. (AC) HCT116 cells were treated with Dox (100 nM) for 4 days in the presence of the indicated doses of Amlexanox or TTM. Then, (A) Cell surface-bound IL-1α were analyzed by FACS (n=3). (B) Cell lysates were subjected to western blot analysis for the indicated proteins. (C) mRNA levels of some SASP genes were analyzed by real-time qPCR (n=3). (D) ER:Ras IMR90 cells were given 4-OHT for toal 6 days in the presence of the indicated doses of Amlexanox; fresh medium with 4-OHT and Amlexanox was changed every other day. Cell lysates were then subjected to Western blot analysis for the indicated proteins. (E–G) HCT116 cells were treated without or with Dox (100 nM) and the indicated doses of TTM for 4 days. Then, cells were collected, washed with PBS for twice, and, (E) Analyzed the intracellular Cu2+ concentration by ICP-MS (n=3). (F) Cell lysates were subjected to western blot analysis for the indicated proteins. (G) mRNA levels of some SASP genes were analyzed by real-time qPCR (n=3). (H) ER:Ras IMR90 cells were treated with TTM as in (D), then the indicated proteins were analyzed by Western blot. Three independent experiments were performed and analyzed. Error bars represent means ± SD (n = 3) *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001 in (A), (C), (E), and (G).