Cell counting kit 8 cck 8

Letrozole Tablets (2.5°mg/piece) were purchased from Hengrui Medicine Co., Ltd. (Jiangsu, China). ELISA kits were purchased from Sinobestbio (Shanghai, China). Primary antibody TLR4 (1:1000, GB11519), p65 (1:1000, GB11997), phosphorylated (p)-p65 (1:000, GB13025-1), Akt (1:1000, GB111114), GAPDH (1:2000, GB11002), and horseradish peroxidase (HRP)-conjugated secondary antibody (1:3000, GB23303) were purchased from Servicebio (Wuhan, China). Primary antibody PI3K (1:1000, A18640), p-PI3K (1:1000, AP0854), and p-Akt (1:1000, AP0637) were purchased from ABclonal (Wuhan, China). RIPA lysis buffer were purchased from Boster (Wuhan, China). RNAprep pure Tissue/Cell Kit, FastKing RT Kit, and SuperReal PreMix Color (SYBR green) were purchased from TIANGEN (Beijing, China). Dulbecco’s modified Eagle's medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin were purchased from Gibco (USA). NO assay kit and cell counting kit-8 (CCK-8) were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). Mag-Bind Soil DNA Kit was purchased from Omega Bio-Tek (United States).

Cell viability was measured using the Cell Counting Kit-8 (CCK-8) (Elabscience, Wuhan, China). Cells were seeded in a 96-well plate and incubated with different concentrations of asarinin for 24 h. After washing the cells with PBS, CCK-8 reagent was added and the cells were incubated at 37 °C for 30 min. The absorbance of each well was measured at a wavelength of 450 nm.

The Cell Counting Kit-8 (CCK-8) (Elabscience, China) assay was used to analyze the proliferation of glioma cells. U87 cells were seeded at a density of 5 × 10³ cells/well in 24-well plates. The cells were incubated in medium containing 40 μmol/L luteolin, quercetin, kaempferol or stigmasterol. The control group was treated with an equal volume of the solvent (DMSO) in the culture medium. After 24 h, glioma cells were then analyzed by CCK-8 assay according to the manufacturer’s instructions. Briefly, 50 μL CCK-8 solution was added to each well. After incubating at 37 °C for 4 h, the absorbance value at 450 nm was measured.