Fig. 3: The nutrient stress metabolome reveals glucose starvation causes disulfidptosis mediated by cystine influx via xCT.
From: The atypical E3 ligase HOIL-1 safeguards the ribosome during cellular stress
a, A schematic of the experimental design examining the metabolome in AC16 cells in full medium or after 3 h or 24 h glucose starvation. b, PLS-DA of the metabolomics dataset. c, A volcano plot of targeted metabolites in parent and HOIL-1ΔRBR AC16 cells after 24 h glucose starvation. Significant when −log(P value) ≥1.3. d, SLC7A11 mRNA levels (normalized to TBP) in AC16 cells after 24 h glucose starvation. Data are represented as the fold change over full medium (n = 3 biological replicates). e, Immunoblot analysis of xCT levels in AC16 cells during glucose starvation. f, A schematic of xCT-dependent nutrient and metabolite flux. g, Cell death in AC16 cells treated with erastin (2 µM), sulfasalazine (500 µM) or DMSO vehicle concurrent with 48 h glucose starvation (n = 4 biological replicates). h, Cell death in xCTKO AC16 cells after 48 h glucose starvation (n = 3 biological replicates). Representative immunoblot of xCT KO. i, The measurement of NADP+ and NADPH ratios during glucose starvation in AC16 cells (n = 3 biological replicates). j, Cell death in AC16 cells treated with BSO (500 µM) or DMSO vehicle concurrent with 48 h glucose starvation (n = 3 biological replicates). k, Cell death in AC16 cells treated with GSH ethyl ester (0.5 mM, 1.0 mM), NAC (0.5 mM, 1.0 mM), TCEP (0.5 mM, 1.0 mM) or H2O vehicle concurrent with 48 h glucose starvation (n = 3 biological replicates). Data are presented as mean ± s.e.m. from one representative experiment. P values were determined by moderated t-tests using the limma R package followed by the Benjamini–Hochberg procedure (c), unpaired two-tailed Student’s t-tests (d and i) or two-way ANOVA followed by Dunnett’s (g and k) or Holm–Šídák’s (h and j) post hoc comparisons.
