A cell counting kit 8 cck

A Cell Counting Kit-8 (CCK-8) (Dojindo, Kumamoto, Japan) was used to determine the cell viability and the half maximal inhibitory concentration (IC50) value of SKOV3 and SKOV3-R exposed to DDP. The cell suspension (4000 cells/100 μL per well) was inoculated into a 96-well plate in a 5% CO₂ incubator at 37 °C for 12 h. Each group was incubated with 200 μL of 1640 medium containing different doses of DPP for 48 h, and then was incubated with CCK-8 reagent for 2 h according to the manufacturer’s instructions. The OD values were determined at 450 nm using the microplate reader Varioskan FLASH (Thermo Fisher Scientific).

A Cell Counting Kit-8 (CCK-8; Dojindo, Japan) was used to estimate cell viability as per the protocols of the manufacturer. Briefly, 3000 cells were seeded into each well of 96-well plates, and the cells were treated with anlotinib at various concentrations. Then, viability analysis was carried out with CCK-8 reagent.

A Cell Counting Kit-8 (CCK-8) (Dojindo, Shanghai, China) was used to measure cell proliferation and detect cytotoxicity, according to the manufacturer’s instructions. Briefly, BmN cells were transfected with pIZT/V5-His-mCherry or pIZT/V5-His-mCherry-BmLHA. Cell were collected in 100 μL aliquots at 24, 48 and 72 h. These 100 μL aliquots were added to individual wells in 96-well plates, together with 10 μL of CCK-8, and incubated for 2 h. Absorbance was measured with a spectrophotometer, at a wavelength of 450 nm. There were three biological sample replicates, and each biological sample replicate included three technique replicates.

The human moderately-differentiated gastric cancer cell line SGC-7901 purchased from the Cell Bank of the Academy of Military Medical Science (Beijing, China) and the human immortalized normal gastric mucosal cell line GES-1 obtained from the Beijing Institute of Cancer Research (Beijing, China) were donated by State Key Laboratory of Cancer Biology, the Digestion Department of XiJing Hospital. The SGC-7901 and GES-1 cells were cultured in RPMI1640 medium (HyClone) supplemented with 10% fetal bovine serum (HyClone) and 1% penicillin/streptomycina in a humidified incubator at 37 °C with 5% CO₂. Cetyltrimethylamonium bromide (CTAB) was purchased from Sigma (St. Louis, MO, USA). Sodium borohydride (NaBH₄), Chloroauric acid (HAucl₄) and Ascorbic acid (AA) were purchased from Aladdin. A Cell Counting kit (CCK-8) was purchased from DoJindo (Kumamoto, Japan). The trypan blue, Hoechst 33258, PI Staining Kits, DCFH-DA fluorescence probe were purchased from Beyotime Company (Shanghai, China). Singlet oxygen sensor green reagent (SOSGR) was purchased from Sigma. SH-PEG-NHS was purchased from local supplier.

Cell culture media HyQ M199/EBSS (M199) and fetal bovine serum were provided from HyClone Laboratories (Logan, UT, USA). RSV, t-BHP, dimethylsulfoxide (DMSO) and compound C were purchased from Sigma–Aldrich (St. Louis, MO, USA). The 5,5,‘6,6'-tetrachloro-1,1‘,3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1), 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR) and the antibody against β-actin were purchased from Beyotime Institute of Biotechnology (Beyotime Biotech, Beijing, China). A cell counting kit (CCK-8) was purchased from Dojindo Laboratories (Dojindo Molecular Technologies, Rockville, MD, USA). Antibodies against PGC-1α, Sirt3, Bcl-2, Bax, AIF and Cytc, mtRNAPol, FoxO3A, caspase 3, phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK) and AMPK were obtained from Cell Signaling Technology (Beverly, MA, USA). MitoSOX Red mitochondrial superoxide indicator for live-cell imaging was obtained from Life Technologies (San Diego, CA, USA).