Fig. 1: Spatial resolution of GPX4-mediated tubular cell ferroptosis in I/R-induced AKI.
From: Autophagy of OTUD5 destabilizes GPX4 to confer ferroptosis-dependent kidney injury
Wild-type C57BL/6J mice were subjected to bilateral ischemia/reperfusion (I/R) injury surgery, or a sham operation, and were sacrificed 48 h after the surgery. Injured and normal sham kidneys were harvested for experiments, including single-cell-RNA-sequencing and spatial transcriptomics. a UMAP projection of 10,971 cells from the sham kidney, and 10,398 cells from the injured kidney; cell identity was annotated based on cell type-specific markers (see the “Methods” section). b A bar plot shows the percentage of each cell type out of the total cells in each group. PT proximal tubular, LOH Loop of Henle, CDPC collecting duct principal cell, MAC macrophage, CDIC collecting duct intercalated cell, EC endothelial cell. c A dot plot shows the ferroptosis-associated signaling pathways among the enriched signaling pathways, using GSEA analysis based on the scRNA-seq data of PT cells from I/R-treated and sham kidneys. d Spatial feature plots and violin plots of the ferroptosis signature score in ST spots. e IHC staining and quantification of 4-HNE expression on kidney sections from sham or I/R-treated mice (n = 5); scale bars, 50 μm, a.u.: arbitrary units. f Schematic of GPX4-mediated lipid peroxidation and cell ferroptosis. g Immunoblot and quantification of GPX4 in mouse kidneys (n = 3 per group), primary renal tubular cells (PRTCs), and HK2 cells. h Spatial feature plots and violin plots of Gpx4 in ST spots from sham or I/R-treated mouse kidneys. i Representative images and quantification of cell membrane lipid peroxidation stained by the liperfluo probe (n = 5 independent experiments), scale bars, 50 μm. j Ferroptosis was measured using the fluorescent lipid peroxidation sensor BODIPY™ 581/591 C11 and cell death Dye 7-AAD in HK2 cells treated with or without Fer-1 following H/R induction (n = 5 independent experiments). Data are presented as mean ± s.e.m.; statistical significance was determined using an unpaired two-tailed Student’s t-test.
