Cell proliferation was assessed using the BrdU Cell Proliferation ELISA Kit (Ab126556, Abcam) following the manufacturer’s instructions. Briefly, 6 × 103 cells were plated in a 96-well plate and subsequently the KD was induced. After incubation for 72 h in hypoxia, BrdU was added to the cells for 3 h. Subsequently, cells were subjected to fixation, permeabilization, and DNA denaturation. Next, cells were incubated with anti-BrdU antibody for 1 h, washed and incubated with peroxidase-conjugated secondary antibody. Finally, the colored reaction that indicates cell proliferation was quantified using a spectrophotometer at a wavelength of 450 nm.

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