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. 2023 Apr;193(4):474–492. doi: 10.1016/j.ajpath.2022.12.009

Figure 1.

Figure 1

Syndecan-1 (Syn-1) treatment improves nanomechanics of endothelial glycocalyx (eGC) and cortex (CTX) after enzymatic degradation with heparinase-I (Hep-I). A–C: Histograms showing eGC height (A), eGC stiffness (B), and CTX stiffness (C) of human umbilical vein endothelial cell (HUVEC) monolayers measured via atomic force microscopy nanoindentation technique. Each dot represents the number of repetitions (each showing mean of 25 to 30 cells). D–F: Statistical analysis of fluorescence intensity differences after stimulation: wheat germ agglutinin (WGA)–stained (D), CD138 antibody (AB)–stained (E), and phalloidin– tetramethylrhodamine (TRITC)–stained (F) HUVEC monolayers. G–I: Representative fluorescence images and fluorescence intensity analyses of WGA-stained (G), CD138 antibody–stained (H), and phalloidin-TRITC–stained (I) HUVECs. J–L: Pearson correlation of eGC height versus eGC stiffness versus CTX stiffness after stimulation. M: Quantification of soluble syndecan-1 in culture medium after Hep-I treatment, measured via enzyme-linked immunosorbent assay. Groups: control (CTR), stimulation with cell culture media; Hep-I, stimulation with media + heparinase-I (0.1 U/μL for 60 minutes); Hep-I + recovery (Rec.), stimulation with heparinase-I (0.1 U/μL for 60 minutes) followed by 24 hours recovery time; Hep-I + Rec.+ Syn-1, stimulation with heparinase-I (0.1 U/μL for 60 minutes) followed by 24-hour recovery time + syndecan-1-treatment (8 pmol/L). Data are given as means ± SD (A–F and M). N = 6 (A–F and M). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001 (one-way analysis of variance). Scale bars = 50 μm (G–I). sCD138, soluble CD138.