Figure 6. The inflammasome regulates the senescence secretome.

(a-b) IMR90 cells were infected with a vector that expresses IL-1α or a control and IF of the indicated SASP components performed. Scale bar, 30 μm. (b) Quantification of (a). (c) IL-1α activates a SASP-like response. IMR90 cells were infected with retroviruses expressing RAS^G12V, IL-1α or Inhibin A. When indicated 4 μM TGFBR1 inhibitor II was used. CM was used to probe chemokine and cytokine antibody arrays. (d) Gene set enrichment analysis (GSEA) of IL1R pathway in the gene expression profile of IMR90 cells undergoing OIS (left) and mouse PANIN (right). FDR, false discovery rate; NES, normalized enrichment score. (e) qRT-PCR analysis showing the expression of a set of genes involved on IL1R signalling. Data are one representative experiment out of 2 independent experiments. (f) IB with antibodies against IL-1α and IL-1β in CM collected from IMR90 ER:RAS (RAS) or IMR90 vector cells (Vector) incubated during 7 days with 200 nM 4OHT and 0.5 % FBS. Pro, precursor form; mat, mature form. (g) Activation of the inflammasome during OIS. IMR90 ER:RAS cells (left) and murine models of SSA (centre) and PANIN (right) display increased Caspase 1 activity. Data are mean ± s.d., n = 4, 3, 6 and 8 different samples for control (GI tract), SSA, control (pancreas), and PanIN respectively. (h) IMR90 ER:RAS cells cultured in the presence or absence of 200 nM 4OHT were subjected to IF with antibodies recognizing inflammasome components. A control of IMR90 ER:RAS cells + 4OHT without primary ab (No ab) is shown in the lower row. Scale bar, 10 μm. (i) IMR90 ER:RAS or IMR90 vector cells were cultured in the presence of 200 nM 4OHT and 0.5 % FBS during 7 days with 10 μM Caspase-1 inhibitor or 20 μM IL1R antagonist. After that time, cells were processed and qRT-PCR against different SASP components performed. Data are mean ± s.d., n = 3 independent experiments. Data from this experiment is also presented in Fig S6j.