Cell light edu dna proliferation kit

The 5-ethynyl-2′-deoxyuridine (EdU) assay was performed using a cell light EdU DNA proliferation kit (RiboBio, Suzhou, China). Bovine preadipocytes were seeded in 24-well cell culture plates and transfected at 50% to 60% density with OE-NC, OE-TORC2, siNC, or siTORC2. After 24 h, the cells were incubated with 5-ethynyl-2′-deoxyuridine medium for 2 h. After 2 h incubation, the EdU assay was performed according to the manufacturer’s protocol.

EdU Staining was performed with a Cell-Light EdU DNA Proliferation Kit (RiboBio, Guangzhou, China). A total of 1 × 10⁵ bovine precursor cells were implanted into 24-well cell culture plates, and transfection (bta-miR-376a mimic and mimic NC) was performed when the cells increased to a density of 50–60%. After 48 h, a cell cycle detection kit was used for detection. Finally, fluorescence photography was carried out under an optical microscope.

Cell proliferation was measured using the Cell Counting Kit-8 (CCK8) (Beyotime, Shanghai, China), 5-Ethynyl-2-deoxyuridine (EdU) assay, and cell cycle assay. The CCK8 assay was performed using a CCK8 kit (Bioscience, Shanghai, China) according to the manufacturer’s instructions with a microplate reader (Infinite M200 PRO; Tecan, Switzerland). EdU assays were performed using a Cell-Light EdU DNA proliferation kit (RiboBio, Guangzhou, China) according to the manufacturer’s instructions with EVOSTM Auto 2 (ThermoFisher, Waltham, MA, USA). Cell cycle assay was performed with the Cell Cycle Straining Kit (MultiSciences, Hangzhou, China) according to the manufacturer’s instructions with ModFit LT^TM (version 5.0; Verity Software House, Topsham, ME, USA). Cell apoptosis was measured using Annexin V-FITC Apoptosis Detection Kit according to the manufacturer’s instructions with ModFit LT^TM (version 5.0; Verity Software House, USA).