Figure 1.
Chemical treatment induces a senescent cellular phenotype. Senescence was induced by palbociclib, except for A549 cells where etoposide was used. (A) Representative β-galactosidase staining was performed to mark senescent cells from indicated cell types after chemical induction of senescence. Scale bars indicate 50 µm. Images are representative of at least three independent experiments. (B) Representative immunofluorescence images from indicated cell types stained for Phalloidin (green) and Hoechst (blue). Scale bars indicate 20 µm. n = 2. (C) qPCR was performed to test CDKN1A gene expression in proliferating or senescent cells. Relative mRNA levels were normalized to the respective proliferating controls. Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (D) Proliferation was quantified in the indicated proliferating or senescent cells using a BrdU Assay. Proliferation rate was normalized to the respective proliferating control. Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (E and F) qPCR was performed to test CXCL1 (E) and IL6 (F) gene expression in proliferating or senescent cells. Relative mRNA levels were normalized to the respective proliferating controls. Data are representative of three independent experiments. All values are means ± SEM. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (G and H) CXCL1 (G) and IL6 (H) secretion of the indicated proliferating or senescent cells was quantified in the supernatant by ELISA. Data are representative of at least two independent experiments. All values are means ± SEM. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test.

Chemical treatment induces a senescent cellular phenotype. Senescence was induced by palbociclib, except for A549 cells where etoposide was used. (A) Representative β-galactosidase staining was performed to mark senescent cells from indicated cell types after chemical induction of senescence. Scale bars indicate 50 µm. Images are representative of at least three independent experiments. (B) Representative immunofluorescence images from indicated cell types stained for Phalloidin (green) and Hoechst (blue). Scale bars indicate 20 µm. n = 2. (C) qPCR was performed to test CDKN1A gene expression in proliferating or senescent cells. Relative mRNA levels were normalized to the respective proliferating controls. Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (D) Proliferation was quantified in the indicated proliferating or senescent cells using a BrdU Assay. Proliferation rate was normalized to the respective proliferating control. Data are representative of three independent experiments. All values are means ± SEM. **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (E and F) qPCR was performed to test CXCL1 (E) and IL6 (F) gene expression in proliferating or senescent cells. Relative mRNA levels were normalized to the respective proliferating controls. Data are representative of three independent experiments. All values are means ± SEM. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test. (G and H) CXCL1 (G) and IL6 (H) secretion of the indicated proliferating or senescent cells was quantified in the supernatant by ELISA. Data are representative of at least two independent experiments. All values are means ± SEM. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001. Statistically significant differences were determined by unpaired Student’s t test.

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