One step tdt mediated dutp nick end labeling tunel apoptosis kit

To conduct histological analysis, the tumors were fixed in 4% paraformaldehyde for 24 h. After deparaffinization, the tissue sections (5 mm) were stained with hematoxylin/eosin (H&E). The tumor cell apoptosis was determined using the One-step TdT-mediated dUTP Nick-End Labeling (TUNEL) Apoptosis Kit according to the manufacturer's instructions (Beyotime, China). All sections were examined using an inverted fluorescence microscope. TUNEL-positive cells were shown in green.

Curcumin, bovine serum albumin (BSA), TBHP, and DMSO were obtained from sigma Sigma-Aldrich (St. Louis, MO, USA). ADMSCs were cultured in the serum-free ncMission hMSC Medium (RP02010, Nuwacell Biotechnologies Co., Ltd, China). Fetal bovine serum (FBS), collagenase II, 0.25% trypsin, 100 U/mL penicillin G and 0.1 mg/mL streptomycin were purchased from Gibco (NY, USA). Chondrocyte growth medium Dulbecco’s Modified Eagle Medium F-12 (DMEM/F12) and phosphate buffered saline (PBS) were obtained from Hyclone (Logan, UT, USA). sEV lipid fluorescent dyes DiI was purchased from Thermo Fisher Scientific (MA, USA). Annexin-V-FITC Apoptosis Detection Kit, 4′,6-diamidino-2-phenylindole (DAPI), EdU Cell Proliferation Kit with Alexa Fluor 488, and One Step TdT-mediated dUTP Nick-End Labeling (TUNEL) Apoptosis Kit were purchased from Beyotime Biotechnology (Jiangsu, China). The following monoclone antibodies and secondary antibody were obtained from Abcam (Cambridge, UK): anti-CD63, anti-tsg101, anti-GM130, anti-8-OHdG, anti-collagen II and anti-aggrecan. The primary antibody of cleaved caspase3 was obtained from Cell Signaling Technology (Danvers, MA, USA).