Fig 2: p16, p53 and VEGF expression in patients with CRC. A. Positive p16staining (200x magnification), B. Negative p16staining (200x magnification), C. Positive p53 staining (200x magnification), D. Negative p53 staining (200x magnification), E. Positive VEGF staining (200x magnification), F. Negative VEGF staining (200x magnification).

Fig 3: Chronic obstructive pulmonary disease (COPD) patients show increased telomere-associated DNA damage foci (TAF) in small airway epithelial cells without significant telomere shortening. Explant lung tissue sections from patients with COPD and lung resection specimens from control subjects containing small airway material were analyzed for expression of senescence-associated markers by immunofluorescence in situ hybridization (immuno-FISH) and immunohistochemistry. A: representative images of immuno-FISH staining for γ-histone protein 2A.X (γH2A.X; green) and telomeres (red) in small airway epithelial cells from patients with COPD and controls captured using ×100 oil objective and following Huygens (SVI) deconvolution. Arrows point to γH2A.X foci colocalizing with telomeres (TAF), depicted by associated histograms and shown at higher magnification on the right (images are from one single Z-plane). B: immuno-FISH images of small airway epithelium in patients with COPD and controls color-coded according to number of TAF (blue: low number; red: high number). C: quantitative FISH (Q-FISH) images of small airway epithelium in patients with COPD and controls color-coded according to telomere length (blue: short; red: long). Dot plots represent percentage of cells positive for TAF and mean telomere length for each individual generated by quantifying Z-stacks of at least 50 cells per subject. The horizontal line represents group median. D: representative images of immunostaining for p16 and SIRT1 (brown) in small airway epithelial cells captured using ×40 objective. Arrows point to positive cells. Bar graphs represent the level of positive staining in the airway epithelium quantified using a semiquantitative scoring method. Values are median + range. E: representative image of double immunofluorescence staining for γH2A.X (green) and p16 (yellow) combined with Q-FISH (red), carried out on lung tissue samples from patients with COPD (n = 8) to determine whether TAF and p16 expression colocalizes. Dot plot represents mean number of TAF in both p16-positive and p16-negative cells per individual, with the horizontal line representing group median. AU, arbitrary units. Statistics: Mann-Whitney U-test: *P < 0.05, **P < 0.01.