Fig 1: Exosomal miR-34a-5p sponged PNUTS to induce cardiac senescence. (A–C) Cardiomyocytes were transfected with Ad-PNUTS or Ad-Ctrl as control. Transfection efficiency was determined using quantitative reverse transcription-PCR (qRT-PCR) (A) and Western blot analysis (B and C). *p<0.05, vs Ad-PNUTS in repeated measures analysis of variance (ANOVA), n=6 (qRT-PCR), n=3 (western blot). (D) Cell cycle distribution was analyzed. *p<0.05 vs control; ?p<0.05 vs exosomePD-1 inhibitor+Ad-PNUTS in ?2 analysis, n=6. (E) Percentage of ß-galactosidase (ß-gal)-positive cells. (F) Representative images of senescence-associated ß-galactosidase (SA-ß-gal) staining. Scale bar: 50 µm. (G–I) qRT-PCR analyzed p21, p16, and telomere length mRNA levels. (J) Relative telomerase activity was measured. *p<0.05 vs control; ?p<0.05 vs exosomePD-1 inhibitor+Ad-PNUTS in repeated measures ANOVA, n=6.