The cells were inoculated into a 96 well plate for treatment with different samples. The cells were then processed according to the user manual of Cell Counting Kit-8 (Beyotime, C0038) and the OD values at the wavelength of 450 nm were measured using a microplate reader. Meanwhile, the ATP and AMP levels were determined using ATP assay kit (Beyotime, S0026B) and AMP-GLO TM assay kit (Promega, V5011) according the procedures in the manufacturer's manual. Briefly, around 1 3 10 6 treated cells were washed with cold PBS and then transferred into a 1.5mL centrifugation vail for lysis. The lysate was centrifuged at 10,000 rpm for 5 min, and the supernatant containing ATP and AMP was analyzed with ATP assay kit (Beyotime, S0026B) and AMP-GLOTM assay kit (Promega, V5011).
Zhao Y., Li M., Yao X., Fei Y., Lin Z., Li Z., Cai K., Zhao Y, & Luo Z. (2020). HCAR1/MCT1 Regulates Tumor Ferroptosis through the Lactate-Mediated AMPK-SCD1 Activity and Its Therapeutic Implications. Cell reports, 33(10).
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