Fig. 7: EMCSs shape the responses of the triple-negative breast cancer subtypes to ferroptosis.
a,b, Box plots of the EMCS-enrichment score, indicating the median line, (1.5× interquartile range (whiskers)) (full list) of each sample calculated by ssGSEA across different TNBC subtypes from a: FUSCCTNBC cohort (two-sided Wilcoxon’s test), n = 258 patient sample (n = 102 BLIS, n = 65 IM, n = 62 LAR, n = 29 MES) and from b: TCGA cohort (two-sided Wilcoxon’s test) n = 158 patient samples (n = 67 BLIS, n = 43 IM, n = 20 LAR, n = 28 MES). c,d, Representative immunoblot (c) for PERK, S1R and MFN2 in MDA-MB-157 and Hs578T cells and relative quantification (d) to MDA-MB-157 cells (n = 3 biological replicates). e, A heat map visualizing the mean of the ERMCS-enrichment score (manually curated and full list) and normalized abundances of PEox across different TNBC subtypes (FUSCCTNBC cohort). min, minimum; max, maximum. f, Representative images of cells from sgCTRL MDA-MB-157 cells transiently transfected with AKAP1–mRFP control or 9xL spacer at time 0 (T0) and after RSL3 (0.5 µM, 6 h (T6h)). The images show SYTOX Green staining, bright field (BF) and RFP. Scale bar, 100 µm. g, The per cent of cell death after RSL3 (0.5 µM, 6 h) of sgCTRL MDA-MB-157 cells transiently transfected with AKAP1–mRFP control or 9xL spacer (n = 3 biological replicates). h, Representative images of sgCTRL and sgPERK MDA-MB-157 cells transiently transfected with AKAP1–mRFP control and sgPERK transfected with OMM–ER linker at time 0 and after 12 h (T12h) of RSL3 (0.5 µM). The images show SYTOX Green staining, BF and RFP. Scale bar, 100 µm. i, The per cent of cell death (RSL3 (0.5 µM, 6 h)) in sgCTRL MDA-MB-157 and sgPERK MDA-MB-157 cells transiently transfected with AKAP1–mRFP control and sgPERK with the OMM–ER linker (n = 3 biological replicates). j, The per cent of cell death after RSL3 (3 µM, 24 h) of HCC1143 cells transiently transfected with AKAP1–mRFP control or OMM–ER linker (n = 3 biological replicates). k, The per cent of cell death after RSL3 (1 µM, 24 h) of Hs578T cells transiently transfected with AKAP1–mRFP control or OMM–ER linker (n = 3 biological replicates). In d, g, i, j and k, the quantitative data are the mean ± s.e.m. In d, the statistical significance was determined by a one-sample t-test. In g, j and k, the statistical significance was determined by a two-sided unpaired t-test. In i, the statistical significance was determined by a one-way ANOVA, Tukey post hoc test. n.s., not significant (P > 0.05), *P ≤ 0.05, **P < 0.01, ***P < 0.001.
