Fig. 4: Ferroptotic cell death exerts inflammation, decreases macrophage-mediated fungal killing, and promotes disease progression during candidiasis.

a BM-derived macrophages were treated with 10 μM Fer-1 for 1 h followed by interaction with C. albicans (MOI 1 and 5) for 4 h. PI⁺ (dead) cells were determined by gating on F4/80⁺ cells. N = 3; duplicate. Two-tailed Mann–Whitney Test. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. b Renal tubular epithelial damage determined by specific ⁵¹Cr release. N = 3; triplicate. Two-tailed Mann–Whitney Test Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. c C. albicans killing of macrophages treated with Fer-1. MOI 1. N = 3; triplicate. Two-tailed Mann–Whitney Test. Bar graphs show mean values ± SEM. d TNFα secretion of BM-derived macrophages during C. albicans infection (MOI 1) for the indicated time points. N = 3, duplicate. Two-tailed Mann–Whitney Test. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. e CXCL8 and TNFα secretion of RTECs in the presence and absence of Fer-1 during C. albicans infection. MOI 5. Two-tailed Mann–Whitney Test. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. f Lactate dehydrogenase (LDH) release of RSL3 (0.5 and 1 μM) treated BM-macrophages. Mann–Whitney Test. N = 3 in duplicate. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. g TNFα release of RSL3-treated or C. albicans-infected macrophages (MOI 1) in the presence or absence of Fer-1 (10 μM). 12 h. Two-tailed Mann–Whitney Test. N = 3; duplicate. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. h Survival of WT mice treated daily with 10 mg/kg Fer-1 or vehicle control. Inoculum 1.25 × 10⁵ C. albicans. N = 5; two independent experiments per inoculum. Mantel–Cox Log-Rank test. i Survival of WT mice treated daily with 10 mg/kg Fer-1 or vehicle control. Inoculum 2.5 × 10⁵ C. albicans. N = 5; two independent experiments per inoculum. Mantel–Cox Log-Rank test. j Renal fungal burden after 3 days of infection. N = 5; combined data of two independent experiments. Two-tailed Mann–Whitney Test. k Serum NGAL and l TREM1 of infected mice after 3 days of infection. N = 5; combined data of two independent experiments. Mann–Whitney Test. m Levels of TNFα, IL-1β, and IL-6 in infected kidneys after 3 days of infection. N = 5; combined data of two independent experiments. Two-tailed Mann–Whitney Test. Results are median ± SEM.