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. 2021 Mar 4;10:e62678. doi: 10.7554/eLife.62678

Figure 4. Serpinh1 expression is increased by aging and obesity and repressed by exercise training.

(A) A Venn diagram showing the overlap of differentially expressed genes between the experiments. Four genes were identified to be significantly affected by aging, obesity, and exercise (Serpinh1, Vwa1, Mest, and Fhl3). (B) Bar plot showing the expression pattern of these four genes. In panels (A and B), the up- and downregulated genes with the false discovery rate (FDR; Benjamini–Hochberg adjusted p-value) threshold of 0.05 were considered to be significant (N = 3–4 male mice/group). (C) qPCR validation of Serpinh1 and Vwa1 normalized to Hprt1 (N = 4–6 male mice/group). (D) In silico secretome analysis of the identified genes. (E–G) Representative immunofluorescent and immunohistochemistry images showing the expression of SERPINH1 in human endothelial cell (EC) and human heart samples. Red arrowhead in the bottom panel F indicates the expression in large vessels and ‘L’ indicates vessel lumen. White arrowheads in the panel G denote the co-expression of SERPINH1 and CDH5 in coronary vessels (yellow signal). (H–K) mRNA expression of Serpinh1, Vwa1, Vim, and Tgfbr2 in the cardiac ECs of sedentary and exercise trained aged mice (N = 4–5 female mice/group). Scale bar 100 μm. Data is presented as mean ± SEM. Student’s t-test was used, *p<0.05, **p<0.01, ***p<0.001.

Figure 4—source data 1. Source data for Figure 4B, C, H, I, J and F.

Figure 4.

Figure 4—figure supplement 1. Expression of Serpinh1 in different cardiac cell types and in the human heart.

Figure 4—figure supplement 1.

A tSNE plot showing (A) cardiac cell types in the adult mouse heart. (B) Violin plots showing the levels of SerpinH1 transcripts in fibroblasts, myofibroblasts, smooth muscle cells, endothelial cells (ECs), endocardial cells, cardiac muscle cells, and leucocytes (each black dot denotes a single cell). (C) tSNE plot showing the expression of Serpinh1 and EndMT genes in the EC cluster (cells within the red circle) and endocardial cluster (cells within the green circle). Illustrations in the panels A–C were analyzed and acquired from a publicly available single-cell database Tabula muris: https://tabula-muris.ds.czbiohub.org/. (D) Representative longitudinal IHC image of human heart demonstrating strong SERPINH1 expression in interstitial cells (fibroblasts, ECs) and weak staining within cardiomyocytes. Scale bar 100 μm.
Figure 4—figure supplement 2. Serpinh1 expression in different subsets of cardiac endothelial cell (EC).

Figure 4—figure supplement 2.

(A–D) A t-SNE plot showing the blood EC (activated EC (Apln+) highlighted within red circle), lymphatic EC (lyve1+ Prox1+), and endocardial EC (Npr3+). (E) Pdgfrb expression in cardiac EC population. (F) Serpinh1 is expressed in all EC populations, and especially highly expressed in Apln+ activated EC (cell cluster within red circle), which also has increased expression of mesenchymal genes (Tagln2, Vim, Smtn, and Cd44). Data in this figure were obtained and analyzed from publicly available EC atlas (https://endotheliomics.shinyapps.io/ec_atlas/).
Figure 4—figure supplement 3. SERPINH1 RNA levels in the human arterial and venous endothelial cells.

Figure 4—figure supplement 3.

RNA expression of SERPINH1 in human arterial and venous endothelial cells in the indicated organs organs. Data obtained by analysing the publicly available datasets (E-GEOD-43475 in the EndoDB database; https://endotheliomics.shinyapps.io/endodb/).