Fig. 5: VICs show pro-inflammatory features during hyperlipidemia. | Nature Communications

Fig. 5: VICs show pro-inflammatory features during hyperlipidemia.

From: Single-cell transcriptomics reveal cellular diversity of aortic valve and the immunomodulation by PPARγ during hyperlipidemia

Fig. 5

a UMAP plot of 1,732 VICs colored by clusters and mouse models as indicated. b Absolute cell numbers (left) and relative proportions (right) of the VIC subsets from each mouse model. c Average expression map of representative genes in each cell cluster. Color depends on the mouse model, and size represents the fraction of cells with the expression value of each gene for each group. d, e Representative gene expression of VIC subclusters. UMAP plot of gene expression (gray to blue) (d) and RNA in situ hybridization (e). Scale bar: 50 μm. f Boxplot of average expression level of genes related to myofibroblast activation (left) and calcification (right) (Gene list in Supplementary Data 2) (n = 981 cells for C57BL/6J; 470 for Apoe−/−; 281 for Ldlr−/−). Each box depicts the IQR and median of each score, whiskers indicate 1.5 times the IQR. p, two-sided T-test p-value. g, h Expression map of monocyte chemoattractant genes. The average expression of genes for VICs (purple to yellow) in each mouse model was scaled by z-transformation and displayed on a scale of at least−2.5 (g) and RNA in situ hybridization of Csf1 and Cx3cl1 (h). Scale bar: 50 μm (left), 20 μm (right). Arrowheads indicate Csf1 (green) and Cx3cl1 (red) signal. i Enrichment plot of significant gene ontology (GO) terms related to inflammation. Genes were ranked by the fold change between knockout and wild-type models for VICs. j Transwell migration assay for evaluating monocyte chemotactic levels of VICs cultured with/without LDL or oxLDL. (n = 4). Samples without VIC were used for control. Size of field: 1272.79 μm2. Scale bar: 150 μm. k Adhesion assay of monocytes to ex vivo cultured aortic valves with/without LDL or oxLDL (n = 6 for non-treated, n = 7 for LDL, n = 6 for oxLDL). Scale bar: 200 μm. Dashed line, outline of valve leaflets. Image data are representative of three independent experiments unless otherwise stated. For (j) and (k), Kruskal–Wallis test with post-hoc Dunn’s test was used, and data are presented as mean ± SD.

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