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. 2019 Mar 27;18(3):e12946. doi: 10.1111/acel.12946

Figure 4.

Figure 4

Caspase‐5 is required for the IL‐1α‐dependent SASP. (a–c) Representative images and quantification of senescence‐associated beta‐galactosidase (SAβG) (a), proliferation by BrdU (b) and senescence‐associated heterochromatic foci (SAHF) (c) in growing (Grow.) and senescent (Sen.) IMR‐90 cells (n.b. b, c are the same field of view). (d–g) Caspase‐5 expression by RNA‐Seq (d), cell surface IL‐1α by flow cytometry (e), cleaved IL‐1α in the conditioned media by ELISA (f) and level of cell death (g) in growing and senescent IMR‐90 cells. (h) ELISA data showing the level of IL‐6, ±neutralizing IL‐1α antibody (α pAb) or IL‐1RA (RA) in the conditioned media of growing and senescent IMR‐90 cells. (i) qPCR data showing relative expression of CASP5 in senescent IMR‐90 cells after transfection of control‐ (Ctrl) or CASP5‐targeted siRNAs. (j–o) Cell surface IL‐1α by flow cytometry (j), or total cleaved IL‐1α (k), non‐calpain‐cleaved IL‐1α (l), IL‐6 (m), IL‐8 (n) and MCP‐1 (o) by ELISA in the conditioned media of senescent IMR‐90 cells after transfection of control‐ or CASP5‐targeted siRNA. Data represent mean ± SEM of n = 3, n = 4 (n, o), n = 6 (d); p = *≤0.05, **≤0.01, ***≤0.001, ****≤0.0001; ns = not significant. Scale bars = 100µm (a), 15 µm (b, c)