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Cells were harvested for assay 48?h after transfection and luciferase activity was measured using the Luciferase reporter system kit (Promega, Madison, WI, USA). Cells were exposed for 2?h to 150?��m H2O2 and the SA-��-gal activity was assessed with SA-��-gal Staining kit (CDK, Englewood, CO, USA) based on the method [http://www.selleckchem.com/products/bmn-673.html Talazoparib nmr] originally described by Oh et?al. (16). Intracellular ROS level was detected using detection kit (Beyotime, Haimen, China). The fluorescent signal was detected with a Fluorescence microplate reader (Thermo Spectronic, Madison, WI, USA) (Ex 488?nm/Em 525?nm). The primer sequence and size of PCR products of target genes are listed in Table?S1. GAPDH primers were used as internal RNA loading and amplification controls. Primary antibodies against ��-catenin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), FoxO3a (Cell Signaling Technology, Irvine, CA, USA), SMP30 (Santa Cruz Biotechnology), Tbx3 (Santa Cruz Biotechnology), p16INK4a (Santa Cruz Biotechnology) and GAPDH (Santa Cruz Biotechnology) were used in western blot analysis. All analyses were performed with SPSS 13.0 (SPSS Corporation, Chicago, CA, USA). Statistical significance of multiple treatments was determined by analysis of variance (ANOVA) test. A P?[https://en.wikipedia.org/wiki/CAPNS1 CAPNS1] (P?[http://www.selleckchem.com/products/s-gsk1349572.html Dolutegravir] To explore the role of ��-catenin on cellular senescence induced by H2O2, NHSFs-vector and NHSFs-��-catenin were treated with H2O2. As expected, after H2O2 treatment, NHSFs-vector became enlarged and irregular in shape and the percentage of SA-��-gal positive cells increased (Fig.?1; P?