Estrogen-Related Receptor Alpha Confers Methotrexate Resistance via Attenuation of Reactive Oxygen Species Production and P53 Mediated Apoptosis in Osteosarcoma Cells

Figure 2

MTX induces cell death by increasing ROS production in osteosarcoma cells. ((a)-(b)) U2OS cells were treated with 1 μM MTX for 24 hours; ROS production was analyzed by DCFH-DA staining using microreader and fluorescent microscope. (c) U2OS cells were pretreated with 25 mM NAC for 2 hours followed by treatment with 1 μM MTX for 24 hours. ROS production was analyzed by DCFH-DA staining using microreader and fluorescent microscope. Data are expressed as mean ± SD (𝑛=3) and analyzed using Student’s 𝑡-test. Asterisks indicate significant differences: ^∗𝑃<0.05, ^∗∗𝑃<0.01.

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