Senescence β galactosidase β gal staining kit

Sodium hydrogen sulfide (NaHS), the anti-CSE antibody, PD98059 (an inhibitor of ERK), LY294002 (a PI3K inhibitor), 5-hydroxydecanoate (5-HD, a mK_ATP inhibitor), Chelerythrine chloride (Che, a PKC inhibitor) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). The primary antibodies for anti-PKC-ε, anti-cleaved caspase-3 and -9, Bcl-2, cytochrome c (Cyt c), Na⁺/K⁺-ATPase, cyclin D1, p21^Cip/WAF-1 and GAPDH were from Santa Cruz (Bergheimer, Germany). Hoechst 33342, JC-1 kit and Calcein-AM were also from Santa Cruz. The anti-ERK1/2 and PI3K-Akt-GSK-3β antibodies were obtained from Cell Signaling Technology (Danvers, USA). Senescence β-galactosidase (β-gal) staining kit was purchased from Beyotime Institute of Biotechnology (Shanghai, China). Rat advanced glycation end products (AGEs) ELISA kit was purchased from Proteintech Group (Wuhan, China). All other chemicals were from Sigma or Santa Cruz.

We obtained the cell culture reagents from Invitrogen (Carlsbad, CA, USA). We purchased D-galactose (D-gal), N-acetyl-L-cysteine (NAC, ROS scavenger) from Sigma-Aldrich (St. Louis, MO, USA), and MCC950 (NLRP3 inflammasome inhibitor), nigericin (NLRP3 inflammasome activator) from MedChem Express (Monmouth Junction, NJ, USA). The senescence β-galactosidase (β-gal) Staining Kit and DCFH-DA probe were purchased from Beyotime (Shanghai, China). The CellEvent™ Senescence Green Flow Cytometry Assay Kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Primary antibodies including P53 (1:1000, Cell Signal Technology, Danvers, MA, USA), P21(1:1000, Cell Signal Technology), β-actin(1:1000, Cell Signal Technology), NLRP3 (1:1000, Adipogen, San Diego, CA, USA) and ASC (1:1000, Abcam, UK) were purchased from the above accompanied. The IL-1β, IL-18 and LDH kits were purchased from Solarbio Technology (Beijing, China).