Extended Data Fig. 2: OGT regulates ferroptosis by controlling the transcription of SLC7A11, related to Fig. 2.
From: A ROS-mediated oxidation-O-GlcNAcylation cascade governs ferroptosis
a-c, The protein levels of SLC7A11 were analysed in PLC/PRF/5 cells following OGT knockdown (a), treatment with OSMI-1 for 24 h as indicated (b), and overexpression OGT (c). Immunoblotting was performed using the specified antibodies (n = 3 experiments). d. e, Endogenous OGT was depleted in PLC/PRF/5 (d) and Huh7 (e) cells using shRNA, and cystine uptake levels were subsequently measured. n = 3 independent experiments. f. g, Endogenous OGT was depleted in PLC/PRF/5 (f) and Huh7 (g) cells using shRNA, and cellular GSH levels were subsequently measured. n = 3 independent experiments. h, Endogenous OGT was depleted in Huh7 cells, treated with the indicated reagents, and lipid peroxidation levels were assessed by flow cytometry. n = 3 independent experiments. i. j, Endogenous OGT was depleted in PLC/PRF/5 (i) and Huh7 (j) cells using shRNA. The cells were treated with increasing concentrations of erastin for 24 h, and cell viability was subsequently measured. Data are presented as mean ± SEM. Two-way ANOVA was used. n = 3 independent experiments. k-m, OGT was stably depleted in PLC/PRF/5 cells by shRNA and stably transfected with SLC7A11. SLC7A11 and OGT protein levels in the indicated cells were determined by Immunoblotting (k). Cellular Cystine uptake (l). and GSH levels (m) in the indicated cells were examined. n = 3 independent repeats. d,e,f,g,h,l,m, Data are presented as the mean ± SEM; two-tailed Student’s t-test; ns, not significant.
