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oai:figshare.com:article/34026594 2026-09-29T17:32:45Z category_4 category_8 category_12 category_13 category_14 category_272 category_46 category_734 category_106 category_132 category_134 portal_5 item_type_1 month_year_09_2026

Recombinant Sph protein induces ferroptotic cell death, which is counteracted by ferroptosis inhibitors. Yunfei Wei (9412265) Xinyu Zhao (102214) Ziyi Yin (4117480) Yanmin Zhang (391964) Yaxin Zhai (2562517) Baihe Zhan (25138380) Yeling Wang (371383) Jiakang He (16829817) Jianfeng Wang (232744) Xuming Deng (158166) Lei Xu (78676) Haihua Feng (198813) Biochemistry Microbiology Cell Biology Genetics Molecular Biology Environmental Sciences not elsewhere classified Immunology Biological Sciences not elsewhere classified Science Policy Infectious Diseases Virology mitochondrial contact site dysregulated inflammatory response disrupt mammary tissues potential therapeutic target key virulence determinant mastitis remain elusive dependent ferroptosis contribute >- induced mastitis >- induced ferroptosis sting signaling pathway mediated mtdna release cgas ), leading staphylococcus aureus </ promote gpx4 degradation aureus </ virulence factors sting axis potential role lysosome pathway underlying mechanisms thereby eliciting sphingomyelin phosphodiesterase organizing system micos ). major pathogen diverse arsenal cyclic gmp core component amp synthase >; however (A) LDH release from MAC-T cells treated with the indicated concentrations of recombinant Sph for 6, 12, or 24 h under continuous incubation at 37°C (n = 5). (B) LDH release from MAC-T cells treated with recombinant Sph under the same conditions, followed by an additional 1-h incubation at 4°C (n = 5). (C) Representative immunoblots showing SLC7A11 and GPX4 expression in MAC-T cells treated with recombinant Sph (4 or 32 μg/mL) for 6 h, with corresponding quantitative analyses (n = 3). (D) Schematic diagram of the experimental protocol for ferroptosis inhibitor intervention in recombinant Sph-treated MAC-T cells. (E-I) LDH release from MAC-T cells treated with recombinant Sph (4, 16, or 32 μg/mL) in the presence of the indicated concentrations (0-30 μM) of ferrostatin-1 (E), zileuton (F), troglitazone (G), TEMPO (H), or deferiprone (I) (n = 3). ##P < 0.01 versus the control group; **P < 0.01 versus the Sph-treated group; ns, not significant. (J) Representative bright-field images of MAC-T cells following the indicated treatments. (K-N) Representative fluorescence images and (O-R) corresponding quantitative analyses of intracellular ROS (DCFH-DA; K, O), Fe2+ accumulation (L, P), lipid peroxidation (C11 BODIPY 581/591; M, Q), and mitochondrial membrane potential (JC-10; N, R) in MAC-T cells treated with recombinant Sph (4 or 32 μg/mL) with or without deferiprone (30 μM) for 6 h (n = 3). Scale bars, 200 μm. Data are presented as mean ± SD. *P < 0.05; **P < 0.01; ns, not significant. 2026-09-29T18:02:15Z Image Figure 10.1371/journal.ppat.1014644.g003 https://figshare.com/articles/figure/_p_Recombinant_Sph_protein_induces_ferroptotic_cell_death_which_is_counteracted_by_ferroptosis_inhibitors_p_/34026594 CC BY 4.0