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Journal: bioRxiv
Article Title: Regulation of Interstitial Lung Diseases by Pulmonary Endothelial Cells via PLVAP
doi: 10.1101/2024.03.12.584592
Figure Lengend Snippet: A. Dot plot showing ligand-receptor pairs between PLVAP-positive endothelial cells and various immune cell subsets in normal controls and patients with different types of ILD (IPF, cHP, NSIP, sarcoidosis, SSc, myositis). B-C. Validation of the SiO2 mouse model. H&E and Masson staining showed increased inflammation and fibrosis in the lung tissue of SiO2-treated mice. D. Human lung tissue CCL14 lung tissue Immunohistochemical staining. Significantly brown-stained CCL14-positive cells were observed in lung tissue of patients with pulmonary fibrosis and controls. (↑indicates immunohistochemically stained positive cells) E. Integration of single-cell sequencing data from GEO multicenter ILD patients and normal controls. Correlation analysis between the PLVAP + vein ECs ratio and IM/AM ratio showed that PLVAP + vein ECs ratio was positively correlated with IM ratio. F. Integration of single-cell sequencing data from GEO multicenter ILD patients and normal controls. Correlation analysis between the IM ratio and Myofibroblast ratio showed that IM ratio was positively correlated with Myofibroblast ratio. G. UMAP shows the distribution of AUCell scores for each cellular Myofibroblast marker gene set in fibroblasts of lung tissues in the sio2 mouse model. Statistical analysis revealed that the AUCell score was higher in the sio2 mouse model group compared with the control group, and the difference was statistically significant. H. UMAP shows the distribution of AUCell scores for each cellular IM marker gene set in lung tissue macrophages of the sio2 mouse model. Statistical analysis revealed that the AUCell score was higher in the sio2 mouse model group compared with the control group, and the difference was statistically significant. I. Correlation analysis between the AUCell scores of Plvap in ECs and AUCell scores of IM marker genes in Macrophage was positive in Sio2 mouse model lung tissue scRNA. J.Correlation analysis between the AUCell scores of Plvap in ECs and AUCell scores of Myofibroblast marker genes in Fibroblasts in scRNA from lung tissue of the Sio2 mouse model was positive. K. UMAP visualization of Macrophage cells, colored by celltype compartment.Macrophage cells subpopulations including Alveolar macrophages (AM), Interstitial macrophages (IM), mixed cells. L. Dot chart shows the marker of IM, mainly focusing on LGMN, MARCKS, AM (PPARG, SIGLECS, FABP4, GCHFR, FN1, INHBA). M. UMAP plot shows scissor-positive and scissor-negative cells, which are associated with low lung function (Low DLCO) and normal lung function (Normal DLCO), respectively. Red and blue dots correspond to low lung function phenotype and normal lung function phenotype, respectively. N. Bar graph showing the proportion of "DLCO-low phenotype" and "DLCO-normal phenotype" cells in AM and IM, respectively. O-Q. Flow analysis plot demonstrating the proportion of IM and AM macrophages in lung tissue of Plvap EC KO mice and control Plvap +/+ mice after 4 weeks of bleomycin modeling. F4/80 + CD11b + /SIGLECF + CD11c + cells were defined as IM/AM cells, respectively, n=5(** p<0.01, ns p>0.05).
Article Snippet: Anti-CCL14 Antibody (R&D Systems, USA) was added to the sections and incubated overnight at 4°C.
Techniques: Staining, Immunohistochemical staining, Sequencing, Marker