Cell Proliferation Analysis using BrdU Assay

Cell proliferation was assessed using an enzyme-linked immunosorbent assay-based and colorimetric BrdU assay (Roche Diagnostics) as previously described [48 (link), 49 (link)]. MCM10-knockdown or shLacZ control J82 and TCCSUP cells were plated into a 96-well plate at density of 3000 cells per well, and cell proliferation was evaluated at 24, 48, and 72 h. After incubation with BrdU for 3 hours at 37°C under 5% CO2, the labeling medium was removed, followed by fixation and final incubation with anti-BrdU-POD solution. The absorbance of the samples was measured using an ELISA reader (Promega) at 450 nm, with the absorbance at 690 nm as reference.