Journal: Nature Communications

Article Title: Macrophage ferroptosis potentiates GCN2 deficiency induced pulmonary venous arterialization

doi: 10.1038/s41467-025-64035-4

Figure Lengend Snippet: a H&E-stained images of 10X Visium spatial transcriptomics sections from Control ( n = 2 individuals) and PVOD ( n = 1 individual) lung tissues. The two control samples represent the upper and lower halves of the same slide (stitched together). Scale bar = 2 mm. b Spatial mapping of tissue region clusters (Alveoli, Bronchi, Vessel, Unspecified) on spatial transcriptomics spots from Control (left) and PVOD (right) lung samples. c Violin plot showing HMOX1 expression levels across tissue regions in Control and PVOD lung samples. P values were determined via two-sided Wilcoxon rank-sum test. d Violin plots depicting expression of arterial endothelial markers ( KDR, CXCL12 ) and venous related marker ( ACKR1 ) in vessel regions comparing Control and PVOD groups. P values were determined via two-sided Wilcoxon rank-sum test. e Violin plots showing arterial and venous endothelial gene set scores in vessel regions of Control versus PVOD samples. P values were determined via two-sided Wilcoxon rank-sum test. f Volcano plot of differentially expressed genes in vessel regions between PVOD and Control groups. P values were determined via two-sided Wilcoxon rank-sum test with Benjamini–Hochberg correction for multiple testing. Significance thresholds were set at |log2 fold change| > 0.5 and adjusted p -value < 0.05. The top 5 upregulated and top 5 downregulated genes are annotated in the plot. GO biological processes ( g ) and KEGG pathways ( h ) significantly enriched (FDR < 0.05) from upregulated genes in PVOD vessel regions. P values were calculated using the hypergeometric test with Benjamini–Hochberg correction for multiple testing. Ten relevant terms associated with pulmonary vascular disease are shown, ranked by combined score. Dot size represents the percentage of genes in the gene set, and dot color indicates –log10(FDR). i Volcano plot of transcription factor activity differences (z-score normalized AUC scores) between Control and PVOD vessel regions analyzed by the limma method. j Violin plot showing ETS1 AUC scores in Control and PVOD vessel regions. k Violin plot of ETS1 expression in venous endothelial cells from scRNA-seq data comparing Control and PVOD groups. P values were determined via two-sided Wilcoxon rank-sum test. l ETS1 transcription factor binding motif (metacluster_183.1) obtained from the cisTarget motif collection (v10nr_clust). m Spatial distribution of cell type proportions (EC_arterial, EC_venous, Macrophages, Muscular cells, Fibroblasts) inferred by RCTD deconvolution. Color intensity corresponds to the relative abundance of each cell type, with darker colors indicating higher proportions. n Heatmaps showing Pearson correlation between RCTD cell type scores and cell death pathway gene set scores in Alveoli (top) and Vessel (bottom) region of the PVOD lung sample (* P < 0.05, ** P < 0.01, *** P < 0.001). P values are indicated in the figures. Source data are provided as a file.

Article Snippet: Fig. 6 Spatial transcriptomics reveals enhanced venous arterialization and ETS1-mediated gene regulation in PVOD lung vessels. a H&E-stained images of 10X Visium spatial transcriptomics sections from Control ( n = 2 individuals) and PVOD ( n = 1 individual) lung tissues.

Techniques: Staining, Control, Expressing, Marker, Activity Assay, Binding Assay