Fig. 1: Mechanical Stress Increased in the ECs from IPF Patient and Significantly Associated with the PF Development.

A The schematic workflow of this study integrated single-nucleus RNA sequencing with bulk RNA-seq data. B Uniform Manifold Approximation and Projection (UMAP) clustering of cells in snRNA-seq data of IPF patients and normal lung tissues. C The “Scissor” algorithm co-analyzed snRNA and bulk-RNA data, selecting lung function-related cell subpopulations from snRNA-seq data using bulk-RNA lung function data. D Stacked bar chart showed the proportions of Scissor subpopulations in each cell type. E The UMAP plot of endothelial cells (ECs) subpopulations in snRNA-seq data. F UMAP plot displaying ECs of Scissor groups. G Volcano plots depicting pseudobulk differentially expressed genes between FLP ECs of IPF group and FLP ECs of Normal group in snRNA-seq data. P values were calculated using the Wald test. Vertical dashed lines represented log2(fold change) threshold of 1, and horizontal dashed lines represented the P-value threshold of 0.05. UMAP plot (H) and violin plot (I) showed mechanical stress (MS) scores of FLP ECs in IPF (n = 4061 cells) and Normal (n = 1826 cells) groups. J Correlation analysis (Pearson Correlation) between mechanical stress score of ECs and fibrosis score of samples shown with error band representing 95% confidence interval. K The UMAP plot showed scissor-positive and scissor-negative cells associated with low lung function (FLP) and high lung function (FHP). UMAP (L) and violin plot (M) showed MS scores of FLP ECs in IPF (n = 5020 cells) and Normal (n = 1383 cells) groups of five GEO datasets. N Meta-analysis of five GEO datasets showing the MS score of ECs in IPF and Normal groups. Standardized mean difference (SMD) were calculated as Hedges’ g with corresponding 95% confidence intervals. O Correlation analysis (Pearson Correlation) of mechanical stress scores and fibroblast subpopulations in GSE136831 dataset. I, M: Violin plots displayed the mean (yellow diamond) the median (center line), the interquartile range, and the whiskers (1.5 × interquartile range). P value was calculated using the two-tailed unpaired t-test. Source data are provided as a Source Data file.