Cell counting kit 8 cck 8

100-μl cell suspensions were prepared in 96-well plates, 10,000 cells were added to each well, five compound holes were set, and the plate was precultured in the incubator for 24 h. A medium containing hederagenin (PureChem-Standard Co., Ltd., Chengdu, China) was put on the culture plate in place of the fresh media. Twenty-four hours were spent incubating the plate. Cell Counting Kit 8 (CCK-8) (Biosharp, Hefei, China) solution was added to each well after the medium had been changed. For 1-4 hours, the dish was incubated. A microplate analyzer was used to test the absorbance at 450 nm. Cell viability was calculated as [(OD_HED − OD_blank)/(OD₀ − OD_blank)] × 100%, in which OD is optical density and HED is hederagenin. Furthermore, OD_HED represents the absorbance of holes with cells, solutions, and drug solutions; OD_blank represents the absorbance of holes with culture media and solutions but no cells, and OD₀ represents the absorbance of holes with cells and solutions but no drug solutions. The quantity of hederagenin digested in U87 cells was also determined using the equation’s results, using the approximate treatment concentration as the 50% inhibitory concentration (IC50).

Cell viability was determined using the Cell Counting Kit-8 (CCK-8) (Biosharp, China) assay following the manufacturer’s instructions. The cells were seeded at a density of 5,000 cells/well in the 96-well plates. After 24 h, cells were treated with different concentrations of UA. The CCK-8 solution was added to each well, and the cells were incubated at 37°C for another 2 h. The absorbance was measured at 450 nm (A₄₅₀). Each sample was replicated thrice. The cells that stained positive for CCK-8 solution was thought to be viable. The results are presented as a percentage compared with sham group.

Cell viability was determined using a Cell Counting Kit-8 (CCK-8) (Biosharp, Beijing) solution. Cells were seeded at a density of 1 × 10⁴ cells/well in 96-well plates and incubated for 24 to 48 h to observe the cell status. After the cells were subjected to H/R as described above, 10 μl CCK-8 (10 mg/ml) solution was added and incubated for 1 h. Absorbance was detected at 450 nm using a microplate reader.

ICG was purchased from Hangzhou Aoya Biotechnology Co., Ltd. High-glucose Dulbecco’s modified essential medium (DMEM), phosphate-balanced saline (PBS), and fetal bovine serum (FBS) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. The Cell Counting Kit-8 (CCK-8) and 4′,6-diamidino-2-phenylindole (DAPI) staining solution were purchased from Biosharp Co., Ltd. RPMI-1640, penicillin, and streptomycin were purchased from Hyclone (Beijing, China). Deionized (DI) water was used in all experimental processes.

2, 2-diphenyl-1-picrylhydrazyl (DPPH) was purchased from TCI Shanghai, ethylene diamine teraacetic acid (EDTA) and ascorbic acid (VC) were purchased from Sinopharm Chemical Reagent Co. (Shanghai, China); all other chemicals used were analytical grade and bought from local suppliers. Dulbecco's modified Eagle's medium (DMEM) high glucose and fetal bovine serum were purchased from HyClone (Ultah, US) and Gibcol Life Technology (New York). Cell Counting Kit-8 (CCK-8) was purchased from Biosharp Life Sciences (Hefei, China). CAT, SOD, GSH-Px, and MDA assay kits were obtained from Solarbio Science and Technology Co., Ltd. (Beijing, China). Murine macrophage cell line RAW264.7 cells were purchased from the Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Simply P Total RNA Extraction Kit was purchased from BioFlux (Hangzhou, China). Prime ScriptTM RT reagent Kit with gDNA Eraser and TB Green^® Premix Ex Taq™ II were obtained from Takara (Beijing, China).

Powder PVA (MW: 7.2–8.14 × 10⁴ g/mol) was purchased from Yingjia Industrial Development Co., Shanghai, China. Cell counting kit-8 (CCK-8) was obtained from Biosharp, Hefei, China. Phosphate buffered saline (PBS), Penicillin/streptavidin (PS), and Trypsin were purchased from HyClone, South Logan, UT, USA. 4’,6-diamidino-2-phenylindole (DAPI) fluorescent staining solution, Calcein-AM/PI Live Dead Cell Assay Kit, TRITC-Phalloidin and Tris(hydroxymethyl)aminomethane hydrochloride (TRIS-HCI) Buffer were purchased from Biyuntian Biotechnology Co., Shanghai, China. TritonX-100 solution, 3-[(3-Cholamidopropyl)-dimethyl-ammonio]-1-propane sulfonate (CHAPS), Tributyl phosphate, N-Decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate (SB3-10), Amidosulfobetaine-14 (ASB-14) and Benzonase nuclease were obtained from Sigma-Aldrich, St. Louis, MO, USA.