Cell counting kit 8 cck 8 solution

The HASMCs were assigned to the following treatments: i) Blank control with no further treatment (control group); ii) negative control treated with LV-NC; and iii) ADAM33 silencing groups treated with LV-ADAM33-shRNA1. After digesting cells with trypsin, HASMCs with a confluence rate of 90% were prepared into a single cell suspension of 5×10⁴ cells/ml in complete medium. The cells were inoculated into 96 well plates (100 µl/ well, i.e. 5×10³ cells/well) and cultured in 5% CO₂ at 37°C for 24 h. Cell Counting Kit-8 (CCK-8) solution (100 µl of 10%; Beijing Transgen Biotech Co., Ltd.) was added into each well. After incubation for 1 h, the optical density (OD) values at 450 nm of the adherent cell layer from each group were determined by enzyme-labeled instrument.