Fig. 7: ST2+ macrophages-derived osteopontin drives neointimal hyperplasia upon endovascular injury. | Nature Communications

Fig. 7: ST2+ macrophages-derived osteopontin drives neointimal hyperplasia upon endovascular injury.

From: Perivascular mesenchymal cells instruct ST2+ reparative macrophages to promote endovascular injury-induced neointimal hyperplasia in mice

Fig. 7: ST2+ macrophages-derived osteopontin drives neointimal hyperplasia upon endovascular injury.The alternative text for this image may have been generated using AI.

a The experimental design. BMDMs treated by IL-33 (20 ng/mL) for 24 h before additional 24 h fresh medium incubation to prepare CMs. b Representative images and quantification of EdU+ VSMCs co-cultured with the indicated CMs. Scale bar, 100 μm. c Analysis of the putative ligand-receptor communications between arterial macrophages and VSMCs based on the scRNA-seq data of D0 and D7 using CellChat. d Volcano plot depicting the upregulated and downregulated genes in macrophages at D0 and D7. e Violin plots showing Spp1 gene expression of the 7 immune cell populations based on scRNA-seq data. f (Left) Representative flow cytometry gating for quantifying OPN-expressing macrophages and neutrophils at D7. (Right) OPN+ cell percentages and OPN signal intensities in macrophages and neutrophils at D7. n = 6 mice per group. g Representative images of immunofluorescence staining of MAC2 and OPN using the artery sections from mice at D7. Scale bars, 50 μm. h mRNA levels of Spp1 in the artery from mice at D0 (n = 4), D2 (n = 8), D4 (n = 8), D7 (n = 8) and D14 (n = 4). i Plasma OPN protein levels in patients with or without in-stent restenosis (ISR, n = 25; Non-ISR, n = 25). j mRNA levels of Spp1 of the artery from Control (n = 7) and Pb-Il33KO (n = 8) at D7. k mRNA levels of Spp1 in the artery from Control (n = 10) and Mac-Il1rl1KO (n = 7) at D7. l mRNA levels of Spp1 in IL-33-treated (20 ng/mL) BMDMs for 24 h. m BMDMs electro-transfected with siSpp1 followed by IL-33 (20 ng/mL) treatment to prepare CMs. n mRNA levels of the cell proliferation-related genes in indicated CM-exposed VSMCs. o Representative H&E staining images and Intima/Media ratios and lumen diameter/aspect ratios in the artery sections at D28. Mice post-FAI were administrated with the hydrogel containing siSpp1 or siNC locally at the injured sites. n = 6 mice for each group. Scale bars, 50 μm. Data are all shown as the mean ± s.e.m. **p < 0.01 or ***p < 0.001 by unpaired two-tailed Student’s t-test (f,i,j,k,n,o), one-way (b,h) or two-way ANOVA (l). Source data and statistic information are provided as Source data files. Figure 7a,c, m were created in BioRender. Shan, B. (2025) https://BioRender.com/x16x8wd.

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