Fig. 2

Decreased SLC7A11 expression augments the susceptibility of CARD9-deficient MDSCs to ferroptosis during C. tropicalis stimulation. (A and B) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1) for 6 h. The expression of Slc7a11, Slc3a2 and Gpx4 mRNA were detected by qPCR. (C and D) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1, 2, 4) for 24 h. The protein levels of SLC7A11 and GPX4 were measured by western blot. (E-G) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1) for 24 h. The glutamate in medium supernatant, intracellular cystine and GSH were determined by the corresponding assay kit. (H-L) WT and Card9^−/− MDSCs were stimulated with C. tropicalis (MOI = 1), meanwhile, only CARD9-deficient MDSCs were treated with vehicle (DMSO) or ferrostatin-1 (Fer-1, 10 µM). After 24 h, the proportion of dead MDSCs was measured by 7AAD (H). The relative cell viability of MDSCs was detected by CCK-8 kit (I). The Lipid ROS of MDSCs was measured by BODIPY 581/591 C11 staining (J). The protein level of 4HNE was determined by western blot (K). The relative MDA level in MDSCs was detected by MDA Assay Kit (L). Data with error bars are expressed as mean ± SEM, n = 3. Each panel shows at least three independent biological replicates from a typical experiment. Statistical analysis was determined by one-way or 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