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. 2012 Feb;165(3):754–764. doi: 10.1111/j.1476-5381.2011.01607.x

Figure 6.

Figure 6

NADPH oxidase is not involved in the effect of pyridoxine in ox-LDL-treated HUVECs. (A) and (B) Cells were pretreated for 15 min with or without pyridoxine (10−7 mol·L−1), apocynin (10 µmol·L−1) or L-NAME (100 µmol·L−1) followed by incubation with 20 mg·L−1 ox-LDL for another 24 h. Endothelial O2•− production was measured by DHE using flow-cytometry (A) and fluorescence microscope (n = 7, *P < 0.05 vs. control, ##P < 0.01 vs. ox-LDL). (C) Western blot analysis showed the expression of p47phox and β-actin. The expression of p47phox was quantified densitometrically as the ratio of p47phox/β-actin (n = 6). (D) Western blot analysis showed the protein expression levels of p47phox in the control siRNA or p47phox siRNA. (E, F) The combination effect of knockdown of p47phox and pyridoxine (10−7 mol·L−1) or L-NAME (100 µmol·L−1) in ox-LDL-treated HUVECs. Endothelial O2•− production was measured by DHE using flow cytometric (n = 4 **P < 0.01 vs. control siRNA, #P < 0.05 vs. ox-LDL, &&P < 0.01 vs. pyridoxine + ox-LDL) and fluorescence microscope.