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L929 conditioned media which contains the macrophage growth factor M-CSF, was prepared by culturing L929 cells (ATCC) in complete DMEM (Thermo Fisher Scientific, MT10013CV) supplemented with 10% FBS, and 1% penicillin and streptomycin for 10 days at 37C, 5% CO 2

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Treatment restructured gut microbiota by increasing beneficial Lachnospiracea, Clostridium, Lactobacillus , and Bifidobacterium while reducing pathogenic Escherichia

ROS level regulation and cell metabolism both affect ferroptosis, and both processes are influenced by heat stress: when increased iron uptake or decreased storage causes iron overload, free iron generates ROS via the Fenton reaction, directly triggering ferroptosis-related damage