Fig. 3

FosB controlsSlc7a11 transcription in MDSCs during C. tropicalisstimulation. (A) Heat map demonstrating differentially expressed genes in the kidneys from Card9^−/− mice infected with C. tropicalis compared with WT mice. (B) The expression levels of Fosb gene in MDSCs treated with or without C. tropicalis for 6 h from RNA-sequencing analysis. (C) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1) for 6 h. The expression of Fosb mRNA was detected by qPCR. (D) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1) for 24 h. The protein level of FosB was measured by western blot. (E-G) WT MDSCs were transfected with Fosb specific siRNA for 24 h, then the cells were stimulated with or without C. tropicalis (MOI = 1) for 6 h (E and F) or 24 h (G). The expression of FosB and SLC7A11 were detected by qPCR and western blot assays. (H) The logo of FosB-binding site from the JASPAR CORE database (https://jaspar.genereg.net/). (I) One potential FosB-binding site in the Slc7a11 promoter were predicted by using the JASPAR CORE database. (J) WT and Card9^−/− MDSCs were stimulated with or without C. tropicalis (MOI = 1) for 6 h. ChIP-qPCR analysis of the interaction between FosB and Slc7a11 promoter binding site. Data with error bars are expressed as mean ± SEM, (A, n = 6) (B-G and J, n = 3). Each panel shows at least six or three independent biological replicates from a typical experiment. Statistical analysis was determined by the unpaired Student’s t-test and two-way ANOVA with Bonferroni’s multiple comparisons test for post-hoc test. ns (not significant), P > 0.05; *P < 0.05, **P < 0.01, ***P < 0.001