Fig. 4: ATF4 orchestrates disulfidptosis.
From: The atypical E3 ligase HOIL-1 safeguards the ribosome during cellular stress
a, Immunoblot analysis of AC16 cells after glucose starvation for the indicated timepoints. b, ATF4 and NRF2 target genes in AC16 cells measured by RT–qPCR. Data are presented as the fold change over full medium (n = 3 biological replicates). c, Cell death in AC16 cells transfected with ATF4, NRF2 or non-targeting control (NTC) siRNA 48 h before glucose starvation for 48 h (n = 3 biological replicates). d, Immunoblot analysis of AC16 cells transfected with ATF4, NRF2 or NTC siRNA 48 h before glucose starvation for 24 h. e, Immunoblot analysis of 786-O cells after 24 h glucose starvation. Two different HOIL-1ΔRBR clones are shown. f, Cell death in 786-O cells transfected with ATF4 or NTC siRNA 48 h before glucose starvation for 48 h (n = 4 biological replicates). g, Cell death in 786-O cells treated with erastin (2 µM), sulfasalazine (500 µM) or DMSO vehicle concurrent with 48 h glucose starvation (n = 4 biological replicates). h, Immunoblot analysis of AC16 cells after DMSO vehicle, CCCP (5 µM, 25 µM) or glucose starvation for 24 h. i, Immunoblot analysis of AC16 cells after amino acid or glucose starvation for the indicated timepoints. j, A schematic of the cell death pathway. Data are presented as mean ± s.e.m. from one representative experiment. P values were determined by unpaired two-tailed Student’s t-tests followed by Holm–Šídák’s post hoc comparison (b, f and g) or two-way ANOVA followed by Dunnett’s post hoc comparison (c).
