Figure 4. Sirt1 is a target of lncRNA Sirt1-AS. (A)Structure of human Sirt1 gene and location of lncRNA Sirt1-AS. (B) RNA-RNA pull-down assay. (C) Fish assay was performed to observe the location of LncRNA Sirt1-AS in senescence HUVECS. (D) ribonuclease protection assay was performed to detect the duplex formation of lncRNA Sirt1-AS and Sirt1. (E) Actinomycin D assay was used to detect the stability of Sirt1 mRNA. (F) Relative expression of Sirt1. (G) Relative mRNA expression of lncRNA Sirt1-AS. (H) Relative mRNA expression of p53. (I–M) Relative protein expression of Sirt1, p16, p53 and p21. (N) p53 acetylation was detected. Error bars represent SD.*, p< 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.