Extended Data Fig. 7: Chemogenetic H2O2 promotes EC senescence.
a, Schematic diagram of the experimental setting: PECs were transduced with lentiviral vectors encoding D-amino acid oxidase (DAAO) targeted to the cell mitochondria for generation of mitochondrial H2O2 in the presence D-alanine (10 mM) for 72 h. Representative images of SA-β-gal staining demonstrates that DAAO-mediated mitochondrial H2O2 induced cellular senescence in PECs, as shown by increased SA-β-gal-positive cells (%) (n = 6). b,c, Immunoblots (b) and γ-H2A.X immunostaining images (c) reveal a marked increase in DDR following mitochondrial H2O2 production in DAAO-transduced PECs incubated with D-alanine (n = 4–6). Scale bar, 20 and 100 μm. Red circles indicate SA-β-gal-positive cells. Error bars represent SD (b). Data represent triplicated biologically independent experiments. P values were calculated using a two-tailed unpaired Student’s t-test (b). Images created with BioRender.com (a). (*P < 0.05). Source data are provided as a Source Data file.
