Fig. 3

Treatment with anti-ageing drugs decreases markers of senescence. A Schematic illustrating the experimental set-up conducted in P10 to P22 HMFs, passaged weekly. B Multi-parameter analysis of senescence markers. Robust Z scores were calculated for a panel of measures relative to the vehicle control. Colour coding used to illustrate the number of Z scores of the experimental drug value from the respective vehicle control mean. Scores highlighted in blue denote a shift towards a more proliferative phenotype and scores highlighted in yellow denote a shift to a more senescent phenotype, all with a robust Z scores of ± 0.5. White indicates no change. N = 2 for all except for SA-β-gal. C P22 HMFs stained with DAPI (blue) and Cell Mask, p21, p16, IL-6, or nucleolin (red), or SA-β-Gal (blue) following 96-day treatment with 5 nM rapamycin, 10 nM dactolisib/BEZ235, 0.1 nM trametinib, or their respective controls. Size bar, 100 μm