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The hAMY 3 R cells were cultured in growth medium consisting of DMEM (Gibco, #31966-021) supplemented with 10% v/v FBS (Gibco, #16140-071), 1% v/v Penicillin-Streptomycin (Gibco, #15140-122), 0.5 mg/mL Geneticin G418 (Gibco, #10131-027), 0.4 mg/mL Hygromycin (Invitrogen, #10687010) and 250 nM Methotrexate (Sigma, A6770)

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Although exploratory experimental studies currently indicate that ferroptosis is involved in the pathology of DN and can be targeted and intervened pharmacologically, the detailed mechanism still needs to be elucidated