Fig. 5.

UCKL1 knockdown reduces the expression of SLC7A11. (a) Volcano plot of differentially expressed genes (DEG, identified by fold change >2 and q-value < 0.05) in doxycycline induced UCKL1 knockdown RKO cells versus control cells. Among the DEGs, SLC7A11 is the most significantly downregulated gene in UCKL1 knockdown RKO cells. (b) Real-time qPCR result of SLC7A11 mRNA expression level in inducible UCKL1 shRNA transduced HCT116 cells with or without doxycycline treatment. Unpaired two-tailed Student’s t-test was used to compare the results. (c) Western blot result of SLC7A11 protein level in doxycycline-inducible UCKL1 shRNA transduced HCT116 cells treated with or without doxycycline. (d) Western blot results of SLC7A11 protein level in doxycycline induced UCKL1 knockdown HCT116 cells versus control cells after SLC7A11 overexpression. (e) Overexpression of SLC7A11 reversed UCKL1 knockdown-induced cell death in HCT116 cells. One-way ANOVA was used to compare the results. (f) Flow cytometry analysis of C11-BODIPY staining revealed that overexpression of SLC7A11 inhibited lipid peroxidation in UCKL1 knockdown HCT116 cells. Bar graph shows lipid peroxidation level in the indicated cells. One-way ANOVA was used to compare the results. (g) Flow cytometry analysis of DCFH-DA staining revealed that overexpression of SLC7A11 rescued total ROS in UCKL1 knockdown HCT116 cells. Bar graph shows fold change of ROS level in indicated cells. One-way ANOVA was used to compare the results. (h) GSEA revealed that UCKL1 knockdown was associated with lipid storage (right) but not the pyrimidine metabolic process (left and middle).