Fig. 6: IL-23 prevents ferroptosis in macrophages and renal tubular epithelial cell during C. albicans infection.

a BM-derived macrophages were infected with C. albicans (MOI 1) for 4 h in the presence and absence of rmIL-23. Total C11ox fluorescence was quantified. Individual cells (N = 62 cells for Ctrl; N = 60 cells for Ctrl + rmIL-23; N = 79 for Ctrl + Candida; N = 57 for Candida + rmIL-23; combined data of three independent experiments) were identified and total fluorescence was measured using ImageJ. Two-tailed Mann–Whitney Test. Violin plot with median and quartiles. b BM-derived macrophages were treated with rmIL-23 for 1 h followed by interaction with C. albicans (MOI 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. c Macrophages-mediated C. albicans killing in the presence and absence of rmIL-23. MOI 1. N = 3 in triplicate. Bar graphs show mean ± SEM. Two-tailed Mann–Whitney Test. d TNFα secretion of BM-derived macrophages during C. albicans infection (MOI 1) for indicated time points. N = 3, duplicate. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. Two-tailed Mann–Whitney Test. e Renal IL-23 p19 levels during disseminated candidiasis. IL-23 p19 levels were determined in kidney homogenates after indicated time points. N = 6 for each time point. Mean values ± SEM. f LDH release of rhIL-23 (250 μg/ml; 12 and 48 h) treated RTECs followed by infection with C. albicans (MOI 5; 5 h). Two-tailed Mann–Whitney Test. N = 3 in duplicate. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values. g Representative images of C11 oxidation of RTECs incubated for 48 h with rhIL-23 followed by C. albicans infection (MOI 1; 2 h). Scale bar 50 μm. N = 10; two independent experiments. h Quantification of total C11ox mean fluorescence intensity (MFI) per high power filed (HPF). N = 10; two independent experiments. Two-tailed Mann–Whitney Test. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values i CXCL8 and TNFα secretion of RTECs incubated with rhIL-23 during C. albicans infection. MOI 5. N = 3 in duplicate. Two-tailed Mann–Whitney Test. Box-and-whisker plots indicating median, 25th/75th percentiles, and the minimum/maximum values.