The BrdU assay was performed according to a previous study [23 (link)]. U251 and U87 cells were seeded at a density of 1 × 104 cells/well in 96-well plates. After cells reached 80% confluence, the cells were labeled with BrdU (Cat#: ab126556, Abcam, Cambridge, UK) for 12 h. After washing twice with PBS, the cells were permeabilized and incubated with BrdU antibody for 2 h at room temperature. Then, the cells were incubated with anti-mouse antibody (Abcam, Cambridge, UK) for 1 h and subjected to a multimode-plate-reader at OD 450 nm (Thermo Fisher Scientific).
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