Cell counting kit 8 cck 8 assay kit

The viability of BEAS-2B cells was assessed by CCK-8 assay using the Cell Counting Kit-8 (CCK-8) assay kit (MedChemExpress, Monmouth Junction, NJ, USA) following the manufacturer’s instructions as previously described [29 (link),30 (link)]. Briefly, transfected or non-transfected cells were inoculated into 96-well plates. At the indicated time points after treatment, 10 μl of CCK-8 solution was added to each well of the plates and incubated for an additional 3 h. Next, the absorbance was examined at 450 nm. Cell viability = (OD _{experiment group –} OD _{blank group})/(OD_{normoxia group} (0 h) – OD _{blank group} (0 h)) *100%.

Calcium chloride dihydrate (CaCl₂·2H₂O), alendronate sodium trihydrate (NaALN·3H₂O) were purchased from Shanghai Aladdin Bio-Chem Technology Co., LTD (Shanghai, China). CDK7 inhibitor THZ1 and the cell counting kit-8 (CCK8) assay kit were purchased from MedChemExpress (MCE, Shanghai). Dimethyl sulfoxide (DMSO) and penicillin–streptomycin (pen/strep) solution (100 ×) were purchased from Sigma Aldrich (MO, USA). Cell culture medium, trypsin–EDTA and fetal bovine serum (FBS) were provided by Gibco (Guangzhou, China). Annexin V-FITC/PI apoptosis detection kits and mitochondrial membrane potential detection kit were purchased from Beyotime (Shanghai, China). 6-carboxy fluorescein (6-FAM), DAPI dihydrochloride and DiR iodide were purchased from Invitrogen (Carlsbad, CA, USA). D-Luciferin potassium salt and Fluo-4, AM were acquired from Yeasen Biotech Co., Ltd (Shanghai, China). Acridine orange-ethidium bromide (AO/EB) staining kit was purchased from Leagene Biotechnology (Beijing, China). Bcl-2, Bax and cleaved caspase-3 antibodies were bought from Absin Bioscience (Shanghai, China). β-actin and secondary antibodies were purchased from Proteintech (Wuhan, China). BCA protein quantitation kit and enhanced chemiluminescence (ECL) kit were purchased from Perkin-Elmer (Waltham, USA). All other reagents were consistent with previous report [35 (link)].

Cell viability was measured using the Cell Counting Kit‐8 (CCK‐8) assay kit (MedChem Express, USA, #HY-K0301). The cells were incubated with 10 µl CCK‐8 solution for 2 h at 37°C. The absorbance value of each well was measured by the microplate reader (BioTek, Richmond, USA).