Detection of MC proliferation with Cell Counting kit-8 (CCK-8) assay

Cells in each group were collected following 96 h transfection, washed twice with PBS, and seeded in a 96-well plate at a density of 2.5x10⁵/ml. Then, 100 µl culture medium (RPMI-1640 supplemented with 10% FBS) was added to each well (in pentaplicate for each group) and cells were cultured at 37˚C for 0, 12, 48, 72 and 96 h. CCK8 reagent (20 µl; Nanjing KeyGen Biotech Co., Ltd) was then added to each well and cells were incubated at 37˚C for 4 h. Absorbance at 450 nm was then measured using a microplate reader.