Electronic Supplementary Material

Figure S1. NOX1 gene expression in endothelial progenitor cells from hypercholestromic rats or after ox-LDL. A. NOX1 mRNA expression in EPCs from hypercholestromic rats. B. NOX1 protein expression in ox-LDL-treated EPCs.Top, Ratio of optical density between NOX1 and -actin; Bottom, representative images of Western blot results for NOX1 and -actin.All values were expressed as means ±S.E.M. (n = 4per group).

Figure S2. Knockdown of nmMLC20suppresses ox-LDL-induced NOX expression and senescence in endothelial progenitor cellsin vitro. A.Representative images of Western blot results for p-nmMLC20, nmMLC20, NOX2, NOX4 and-actin. B. Ratio of optical density between p-nmMLC20(top) or nmMLC20(bottom) and -actin. C. Ratio of optical density betweenNOX2(top) or NOX4 (bottom) and -actin. D. NOX activity in endothelial progenitor cells. E. H2O2 contents in endothelial progenitor cells. F. Right panel, representative images of -galactosidase staining for endothelial progenitor cells; Left panel, the percentages of -gal positive cells in each group.All values are expressed as mean  SEM (n = 6per group). NOX: NADPH oxidase; +nmMLC20 siRNA: ox-LDL+nmMLC20siRNA;+ siRNA NC: ox-LDL + siRNA negative control. **P 0.01 vs Control; ##P 0.01 vs ox-LDL.

Figure S3. Knockdown of NOX2 or NOX4 attenuates ox-LDL-induced senescence of endothelial progenitor cellsin vitro. A.Representative images of Western blot results for NOX2, NOX4 and-actin. B. Ratio of optical density betweenNOX2 and -actin. C. Ratio of optical density betweenNOX4and -actin. D. NOX activity in endothelial progenitor cells. E. H2O2 contents in endothelial progenitor cells. F. Right panel, representative images of -galactosidase staining for endothelial progenitor cells; Left panel, the percentages of -gal positive cells in each group.All values are expressed as mean  SEM (n = 6per group). NOX: NADPH oxidase; +NOX2 siRNA: ox-LDL+NOX2 siRNA; +NOX4 siRNA: ox-LDL+NOX4 siRNA; + siRNA NC: ox-LDL + siRNA negative control. **P 0.01 vs Control; ##P 0.01 vs ox-LDL.

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