Extended Data Fig. 9: BAI1 suppresses miMOMP and SASP in senescent cells and does not inhibit DNA-induced inflammation.
From: Apoptotic stress causes mtDNA release during senescence and drives the SASP
(a) Percentage cell death following ABT737 + S6 induced cell death at hour 8 post apoptosis induction in presence of vehicle control or BAI1. Data are representative of n = 3 independent experiments. (b) Western blot showing successful CRISPR/Cas9-mediated deletion of BAX/BAK and single BAX and BAK in U2OS cells. HSP60 was used as loading control. Image is representative of two separate blots. (c) Representative image of TOM20 (green) and BAX6A7 (red) in senescent (IR) MRC5 fibroblasts treated with DMSO or BAI1 (scale bar is 25 µm). (d) Percentage of MRC5 fibroblasts containing BAX6A7-positive mitochondria following BAI1 treatment (n = 3 independent experiments). (e) mRNA expression levels of the indicated SASP genes in BAI1-treated senescent MRC5 fibroblasts (n = 3 independent experiments). (f) Heatmap showing secreted levels of IL-6, IL-8 and IP-10 in proliferating and senescent MRC5 fibroblasts treated with different concentrations of BAI1. Values are shown as fold change to proliferating controls. Data are mean of n = 4 independent experiments. (g) Percentage death of vehicle- or BAI1- treated proliferating and senescent cells upon ABT263 treatment at the concentrations indicated. Data are mean of n = 3 technical replicates ± S.E.M. mRNA levels of (h) IL-6, (i) IL-8, and (j) IL-1β in control (EmptyCRISPR) and BAX/BAK-/- MRC5 fibroblasts treated with Herring testes DNA (HT-DNA) with or without BAI1 (n = 3 independent experiments). mRNA expression levels of (k) IL-6, (l) IL-1α, (m) IL-1β, and (n) IL-8 in senescent cells treated with eltrombopagan (EO) (n = 3 independent experiments). Data are mean ± S.E.M. Statistical significance was assessed using one-way ANOVA followed by Tukey’s multiple comparison test (d, f, k-n), two-way ANOVA followed by Sidak’s multiple comparisons test (h-j) or Tukey’s multiple comparison test (a, e, g). For gel source data (b), see Supplementary Fig. 1.
