Fig. 1

CARD9 ablation promotes acute kidney injury and renal ferroptosis during disseminated candidiasis. (A-C) RNA-sequencing analysis was performed with the kidneys of WT and Card9^−/− mice (n = 6 for each group) infected with C. tropicalis. The volcano map showed all the differentially expressed genes (A). KEGG and GO pathway enrichment analysis (B and C). (D) The expression of the indicated genes in the kidneys of WT and Card9^−/− mice infected with C. tropicalis were detected by qPCR. (E) Western blot analysis of SLC7A11 and GPX4 in the kidneys of WT and Card9^−/− mice infected with C. tropicalis. (F) The protein levels of SLC7A11 and GPX4 in infected kidneys were measured by IHC. Scale bar: 50 µM. The percentages of SLC7A11-positive and GPX4-positive were calculated. (G) 4HNE protein expression in infected kidneys was determined by immunofluorescence. 4HNE shown in red. DAPI (blue) was used to stain the nuclei. Scale bar: 50 µM. (H) The relative MDA level in infected kidneys were detected by MDA Assay Kit. (I) The cell death in infected kidneys was determined by TUNEL staining. TUNEL staining shown in green. DAPI (blue) was used to stain the nuclei. Scale bar: 50 µM. Data with error bars are expressed as mean ± SEM, n = 6. Each panel shows six independent biological replicates from a typical experiment. Statistical analysis was determined by two-way ANOVA with Bonferroni’s multiple comparisons test for post-hoc test (D) and the unpaired Student’s t-test (F and H). ns (not significant), P > 0.05; *P < 0.05, **P < 0.01, ***P < 0.001