Fig. 2.

sEVs act as part of the SASP and transfer senescence to co-cultured NHBEs in submerged assays. A Increases in IL6 with addition of DHBE-derived sEVs and B A similar pattern seen with IL8. C–H Increased senescence related gene expression with co-culture of DHBE-derived sEVs; C p16, D p21, E TP53, F Sirt1, and G BTG2 expression increases with addition of DHBE-derived sEVs. Gene normalisation to GAPDH for δδCT and graphs show fold change relative to untreated control group. H Representative images of γH2AX (green) and DRAQ5 (red) staining upon addition of NHBE and DHBE-derived sEVs. I Quantification of γH2AX showing increased DNA damage with DHBE-derived sEV addition. J Representative images of SA-β-Gal (green) and DRAQ5 (red) staining upon addition of NHBE and DHBE-derived sEVs. K Quantification of SA-β-Gal showing increased expression with DHBE-derived sEV addition. N = 6–8. Each colour represents a different donor and experiment. Significant differences between groups shown **p < 0.01; *p < 0.05, Wilcoxon test and Friedmans test