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. 2021 Dec 17;10(12):3563. doi: 10.3390/cells10123563

Figure 2.

Figure 2

Expression profile of EndMT markers in varicose veins. (A) Scatter plots of mRNA fold changes of EndMT-associated genes such as SNAI1, SNAI2, TWIST1, TWIST2, TGFβ1-3, BMP4, SMAD1-5, and SMAD9 in 39 human varicose and 44 control saphenous veins. GAPDH was taken as the endogenous control for quantification. Horizontal lines in scatter plots indicate median values. (B) Representative photomicrographs of immunostaining illustrate that SNAI1/2, TWIST1, TGFβ1, BMP4, pSMAD2, and pSMAD5 proteins were overexpressed in varicose compared to healthy saphenous veins. The areas enclosed by dotted lines show the differential expression of EndMT markers in the neointima of varicose veins. In the control vein, the dotted box indicates the protein expression in intima. (Scale bar 100 µM, magnification 20×). (C) Semiquantitative H score analysis shows overexpression of SNAI1/2, TWIST1, TGFβ1, BMP4, pSMAD2, and pSMAD5 proteins in varicose veins, which was further substantiated with (D) Western blot assays performed using total protein extracted from healthy veins (lanes 1–3) and human varicose veins (lanes 4–6). (E) Bar graph demonstrating densitometry analysis of immunoblots from 31 varicose veins compared to 34 healthy veins indicated significant upregulation of EndMT markers in varicose veins. Values are means ± SD (standard deviation) * indicates p < 0.05 vs. healthy subjects, ** p < 0.01 and **** p < 0.0001.