Cck 8 detection kit

HCT-116 cell viability was evaluated using CCK-8 Detection Kit (Beyotime Institute of Biotechnology, Shanghai, China). HCT-116 cells were seeded into 96-well plates and subjected to indicated treatments. The cells were further maintained in 90 μl of culture medium plus 10 μl CCK-8 reagents per well for 1 h. The optical density at 490 nm (OD 490 nm) in each well was determined by an enzyme immunoassay analyzer (Bio-Tek ELX-800; Winooski, VT, USA).

The CCK-8 detection kit (Beyotime Institute of Biotechnology, Nantong, China) was used to measure cell viability according to the manufacturer’s protocol. Cells were seeded onto 96-well plates (5×10³ cells per well). After 24 h, cells were treated with different concentrations of Glaucocalyxin A. The cells were cultured respectively for 24 and 48 h. Subsequently, CCK-8 solution was added to each well, and incubated at 37 °C for an additional 3 h. The viable cells were counted by absorbance measurements with a monochromator microplate reader at a wavelength of 450 nm. The optical density value was reported as the percentage of cell viability in relation to the control group (set as 100%).

Cell proliferation was determined by a CCK-8 detection kit (C0038; Beyotime, Haimen, China). Briefly, transfected cells were seeded into 96-well plate (5,000/well) and cultured for the indicated times. CCK-8 solution (10 µl) was added to each well at specific time points, and the absorbance at 450 nm was determined by a plate reader. Each experiment was independently repeated for three or five times. For apoptosis assay, the ccRCC cells were collected, stained with the annexin V-conjugated fluorescein isothiocyanate (FITC) and the propidium iodide (PI) (C1062S-2; Beyotime, Haimen, China) following the manufacturer’s instructions, and analyzed using FACScan™ flow cytometer (BD Biosciences).

The 5-ethynyl-2′-deoxyuridine (EdU) cell proliferation kit (RiboBio, Guangzhou, China. Catalog No. C10310) was used to determine cell proliferation. According to the manufacturer's instructions, EdU was infiltrated into the proliferating cells, and the ratio of proliferating cells was calculated by fluorescence microscopy.
Cell proliferation was also determined by Western blot analysis of proliferating cell nuclear antigen (PCNA). PCNA antibody (Cell Signaling Technology, MA, USA. Catalog No. 13110) and GAPDH antibody (Cell Signaling Technology. Catalog No. 2118) were used in the experiment. GAPDH was used as an internal control.
Cell viability was determined using the CCK8 detection kit (Beyotime, Wuhan, China. Catalog No. C0037) according to the reagent manufacturer's instructions. A microplate reader (iMark™ Microplate Absorbance Reader, Bio-Rad, CA, USA) was used to detect the absorbance of the sample at 450 nm (optical density at 450, OD 450). Cell viability was expressed as a percentage.

The CCK-8 detection kit (Beyotime, China) was used to investigate the effects of different compounds on cell viability, as described previously (7 (link)). Briefly, approximately 5 × 10³ GCO cells were seeded into 96-well plates. Cells were treated with compounds at different concentrations for 24 h. Then, 10 μL of CCK-8 solution was added to each well and incubated at 28°C for 4 h, and the absorbance at 450 nm was measured using a microplate reader (Bio-Rad, USA). The untreated cells were considered the positive control, while the wells containing no cells and only culture medium were used as blank controls. Data are presented as the means ± SD of three replicates.

The cytotoxicity of DNase‐I/HSA NMs was measured by CCK‐8. Briefly, MH‐S cells and A549 cells were incubated with different concentrations of NMs (range from 0 to 800 µg mL⁻¹) for 24 h, respectively. After that, the NMs were removed by washing with PBS. For CCK‐8 assay, the CCK8 detection kit (Beyotime) was used according to the manufacturer's protocol.