Journal: Aging Cell

Article Title: Mitigating Pro‐Inflammatory SASP and DAMP With Urolithin A: A Novel Senomorphic Strategy

doi: 10.1111/acel.70237

Figure Lengend Snippet: Doxorubicin treatment induces senescence in human IMR‐90 fibroblasts. (A) Representative images of SA‐βGal‐stained S‐dox and NS with or without the addition of Urolithin A (UA). (B) The proportion of cells staining positive for SA‐βGal in NS and S‐dox cells with and without the UA treatment. Four fields were quantified per well ( n = 3). (C, D) Relative p16 (C) and p21 (D) expression in NS, S‐dox, and RS cells with and without UA treatment, n = 4. (E) IF staining for γ‐H2AX and HMGB1 in NS, S‐dox, and RS cells with or without the addition of UA, 10 days post‐induction. γ‐H2AX—green, HMGB1—red, and DAPI—blue. Scale bar = 30 μm. (F, G) The percentage of cells with two or more γ‐H2AX foci per cell (F) and with nuclear HMGB1 (G). (H–J) Fold change of IL6 (H), IL8 (I), and IL1α (J) gene expression in S‐dox and RS cells relative to NS control. n = 4. All data are presented as mean ± SEM. One‐way ANOVA. ** p < 0.002, *** p < 0.0002 **** p < 0.0001.

Article Snippet: Primary antibodies used included γH2AX mouse antibody (Novus Biologicals, NB100‐74435), pSTING rabbit antibody (Cell Signaling Technology, 19781S) and anti‐HMGB1 rabbit antibody (Abcam, ab18256).

Techniques: Staining, Expressing, Gene Expression, Control