Fig. 3: FSP1 is upregulated in KRASG12D-expressing cells and mediates ferroptosis resistance.
From: Elevated FSP1 protects KRAS-mutated cells from ferroptosis during tumor initiation
a KRAS WT or KRASG12D-expressing cells were subjected to RNA-sequencing. False discovery rate (FDR) [−Log10] is shown for KEGG pathways significantly enriched within the top 1000 genes upregulated in KRASG12D cells. b Hierarchical clustering of fold change (FPKM + 0.01) of ferroptosis KEGG + genes in KRAS WT and KRASG12D-expressing cells. c Levels of FSP1 mRNA were quantified by qPCR in Rasless MEFs expressing WT or KRASG12D d Indicated cells were treated with RSL3 [100 nM] for 5 h and subjected to Western blotting. e, f The indicated cells were subjected to FSP1 or control knockdowns for 48 h and subsequently treated with RSL3 [100 nM] for 24 h. Cell death was determined by flow cytometry and propidium iodide (PI) incorporation. 0 % PI-Incorporation is gated to untreated control. Western blots of representative control lysates are shown. g, h WT and KRASG12D cells stably overexpressing FSP1 were generated and cells were treated with RSL3 [100 nM] alone or in combination with Ferrostatin-1 (Fer-1) [5 µM] for 24 h. Cell death was determined by propidium iodide (PI) uptake and flow cytometry. 0 % PI-Incorporation is gated to control untreated. Western blots of representative control lysates are shown. Data are means ± SEM of three independent experiments in each individual cell line or representative images or histograms were applicable. Two-tailed t-test (c), Two-way ANOVA + Tukey’s multiple comparison test (e, g), ****p < 0.0001, *p < 0.05. Uncropped blots are provided as Original Data file.
