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. 2021 Feb 25;9:616893. doi: 10.3389/fcell.2021.616893

FIGURE 8.

FIGURE 8

Extracellular matrix (ECM) deposition and its tissue-specific patterns after treatment of stromal cells by menstrual blood discharge or peripheral venous blood serum. (A) Representative Western blotting of endometrial stromal cell (EndoSC), dermis stromal cell (DermSC), and adipose tissue stromal cell (AdipoSC) lysates treated with 10% blood serum (BS), menstrual discharge serum (MDS), or fetal bovine serum (FBS) at 96 h of experiment. (B) Densitometry of extracellular matrix (ECM) components in stromal cells treated by serum preparations shown in panel (A). Individual donor data (n = 3) presented as fold change relative to the evaluated protein level in control sample treated with FBS (logarithmic scale); glyceraldehyde 3-phosphate dehydrogenase (GAPDH) loading control used for normalization. Paired t-test was used for analysis of log-transformed data; *p < 0.05. (C) MDS induces anisotropic deposition of extracellular collagen I in stromal cells from the dermis and adipose tissue, but not endometrium. In contrast, stromal cells cultured with BS demonstrated a homogeneous deposition of collagen I and ED-A fibronectin. For EndoSC, response to MDS or BS demonstrates minimal differences and vividly homogeneous deposition of ECM after both treatments. Merged extracellular collagen I and ED-A fibronectin immunofluorescence; nuclei stained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar 100 μm. (D) Immunfluorescence of extracellular collagen I and ED-A fibronectin demonstrated a higher level of colocalization in EndoSC compared to DermSC and AdipoSC independently of serum treatment used in cultures; Pearson’s correlation coefficient for three repeats; one-way ANOVA and Newman–Keuls test were used for statistical analysis; *p < 0.05.