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. 2017 Dec 22;19(1):63–75. doi: 10.1080/15384047.2017.1385675

Figure 3.

Figure 3.

DOXO and IRINO are the strongest inducers of cellular senescence and activate its hallmarks: growth arrest, and an increase in size, granularity and SA-β-gal positivity. HCT116 cells were treated with LONG CHEMO protocol. A. Representative photos showing morphological alterations in OXA-, 5-FU-, DOXO- or IRINO-treated HCT116 cells. Cell nuclei were stained with H33342 dye (blue). Data acquired using fluorescence and transmitted light microscopy (200x magnification) at the end of the 5th cycle of LONG CHEMO protocol. Scale bar – 100μm. B. Evaluation of cell number at several time points after drug treatment. Cell were counted in Bürker's chamber. C. Fraction of granular cells as determined by FSC/SSC analysis, using flow cytometry. D. Representative photos of SA-β-gal staining on untreated and treated cells. Staining was performed on cytospined cells to enable easier quantification of SA-β-gal positive cells. Scale bar – 100μm. Data obtained with transmitted light microscopy (200x magnification) at the end of the 5th cycle of LONG CHEMO protocol. E. Fractions of SA-β-gal+ cells. Each bar represents mean ± SEM, N≥3; statistical significance * – p < 0,05, ** – p < 0,01, *** – p < 0,001 – untreated vs. LONG CHEMO.