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 = 4 per group).
Figure S2. Knockdown of nmMLC20 suppresses ox-LDL-induced NOX expression and
senescence in endothelial progenitor cells in 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
between NOX2 (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 = 6 per group). NOX:
NADPH oxidase; +nmMLC20 siRNA: ox-LDL+nmMLC20 siRNA; + 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 cells in vitro. A. Representative images of Western blot results for
NOX2, NOX4 and -actin. B. Ratio of optical density between NOX2 and -actin. C. Ratio of
optical density between NOX4 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 = 6 per 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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