Figure 6.
Valve population is composed of two subclusters with different gene expression. (A) Subclustering analysis of the valve cluster revealed two valve subclusters, corresponding to the LECs on the upstream sides of the valve leaflets, and LECs on the downstream sides of the valve leaflets. (B) Volcano plot of DEGs between the upstream valve LEC and downstream valve LEC clusters. The horizontal line shows the log2 fold change (FC) threshold set at 0.75. Vertical lines show the significance threshold for adjusted P values set at an FDR of 0.05. (C) Individual UMAP plots showing the expression of DEGs in upstream valve LECs (GJA1, ITGA9, NEO1, CD24) and in downstream valve LECs (GJA4, CLDN11, FOXC2, ANGPT2). (D and E) Confocal analysis of valves present in the upper 200 µm of human dermis confirmed (D) the differential expression of FOXC2 and CD24 by LECs on different sides of the valve leaflet, and (E) the colocalization of CD24 and NEO1 by LECs on the same valve leaflet side. Representative images from n = 2–3 independent experiments (donors) are shown in D and E. Scale bars: 20 µm. (F) Schematic illustration of the structure of the valves and the positioning of the LECs within the valves, based on Saygili Demir et al. (2023). (G) Schematic illustration of bulk RNA-seq of human dermal LECs subjected to laminar shear stress (4 dyn/cm2), oscillatory-like shear stress (laminar shear stress at 4 dyn/cm2 that reverses direction by 180° every 4 s), or static conditions for 48 h. The top 50 DEGs in the laminar and oscillatory conditions are shown in Data S2 and S3. (G) Pearson correlation plot of the bulk RNA-seq described in E. (H) PCA plot of the samples described in G analyzed at three different conditions (static, oscillatory, and laminar). (I) Similarity of LEC clusters upstream and downstream of valves with the bulk RNA-seq of human dermal LECs subjected to static conditions; laminar or oscillatory flow was determined by the correlation of the top 100 DEGs of valve LEC clusters (Data S1) with the bulk RNA-seq dataset. FDR, false discovery rate.

Valve population is composed of two subclusters with different gene expression. (A) Subclustering analysis of the valve cluster revealed two valve subclusters, corresponding to the LECs on the upstream sides of the valve leaflets, and LECs on the downstream sides of the valve leaflets. (B) Volcano plot of DEGs between the upstream valve LEC and downstream valve LEC clusters. The horizontal line shows the log2 fold change (FC) threshold set at 0.75. Vertical lines show the significance threshold for adjusted P values set at an FDR of 0.05. (C) Individual UMAP plots showing the expression of DEGs in upstream valve LECs (GJA1, ITGA9, NEO1, CD24) and in downstream valve LECs (GJA4, CLDN11, FOXC2, ANGPT2). (D and E) Confocal analysis of valves present in the upper 200 µm of human dermis confirmed (D) the differential expression of FOXC2 and CD24 by LECs on different sides of the valve leaflet, and (E) the colocalization of CD24 and NEO1 by LECs on the same valve leaflet side. Representative images from n = 2–3 independent experiments (donors) are shown in D and E. Scale bars: 20 µm. (F) Schematic illustration of the structure of the valves and the positioning of the LECs within the valves, based on Saygili Demir et al. (2023). (G) Schematic illustration of bulk RNA-seq of human dermal LECs subjected to laminar shear stress (4 dyn/cm2), oscillatory-like shear stress (laminar shear stress at 4 dyn/cm2 that reverses direction by 180° every 4 s), or static conditions for 48 h. The top 50 DEGs in the laminar and oscillatory conditions are shown in Data S2 and S3. (G) Pearson correlation plot of the bulk RNA-seq described in E. (H) PCA plot of the samples described in G analyzed at three different conditions (static, oscillatory, and laminar). (I) Similarity of LEC clusters upstream and downstream of valves with the bulk RNA-seq of human dermal LECs subjected to static conditions; laminar or oscillatory flow was determined by the correlation of the top 100 DEGs of valve LEC clusters (Data S1) with the bulk RNA-seq dataset. FDR, false discovery rate.

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