Fig. 5

EGCG-EVs rescue hADMSC from serum-starvation-induced senescence. hADMSC were incubated for 24 h in complete media (CM), serum-deprived basal media (BM), EVs or EGCG-EVs at a ratio Cell:EVs of 1:0.5. hADMSC were collected for protein and total RNA as described in the Methods section. A Immunoblotting detection of the senescence biomarker p21 and of the loading control tubulin from control hADMSC lysates, treated with CM, BM, or the respective EVs. Immunoblotting is representative of three independent experiments. B Confocal microscopy of hADMSC treated for 48 h at a Cell: EVs ratio of 1:2. The nucleus was stained with DAPI (red), and the expression of the senescence-associated β-galactosidase (β-gal) marker is coloured in green. One out of three experiments is presented. C Histograms showing the percent of positive β-gal cells obtained upon 48 h of treatment. D Gene expression of other senescence markers modulated in hADMSC by EGCG-EVs (black bars) compared with the expression level of the genes in cells incubated with EVs (white bars), using as cut-off a log2 FC ≥ 2 and quantified by qPCR using the Human Senescence RT2-Profiler gene array kit. The percent of positive β-gal cells/field was calculated using the following equation: (number of positive cells /total of cells)*100. The Kruskall-Wallis test determined statistically significant differences, showing a * p < 0.05