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. 2019 Jun 25;20(12):3107. doi: 10.3390/ijms20123107

Figure 5.

Figure 5

Expression of MΦ soluble endoglin induced by MMP-12. (A) Effect of the MMP-12 specific inhibitor MMP-408 on the release of soluble endoglin during in vitro macrophage polarization of human monocytes. (B–D) Effect of MMP-12 silencing in GM-MΦ. GM-MΦ were transfected with siRNA targeting MMP-12 or negative control (siC). After 48 h, MMP-12 mRNA (B) and protein (C) levels were determined by qRT-PCR and ELISA, respectively. Soluble endoglin levels were also measured in GM-MΦ culture supernatants (D). (E,F) Recombinant expression of MMP-12 releases soluble endoglin from membrane bound endoglin. COS-7 cells were co-transfected with endoglin (ENG), GFP, and MMP-12, as indicated. Transfection efficiency was visualized by fluorescence microscopy of GFP-expressing cells (E, upper panel) and by Western blot analysis (E, lower panel). A negative control with an empty vector (Φ) is included. Soluble endoglin levels were measured by ELISA in culture supernatants from transfected COS-7 cells. Statistical analysis was calculated using the paired two-tailed Student t test (n = 4), and data are presented as mean ± SD. (*p < 0.05; **p < 0.01).