Skip to main content
. 2020 Jul 13;36:101638. doi: 10.1016/j.redox.2020.101638

Fig. 2.

Fig. 2

NOX4 knockdown decreases ROS production in CLP-induced septic mice. Sepsis was induced by CLP in wild-type (WT) mice and mice deficient in NOX2 or p47phox, or mice transfected of control siRNA, NOX1 or NOX4 siRNA. Mouse lung tissues were freshly harvested at 16 h post CLP for DHE fluorescent imaging, or at 1, 2, 4, 6 h post CLP for Western blotting analyses. (A): Representative images of DHE fluorescent imaging of lung tissue sections in sham group, CLP groups treated with control siRNA, NOX1 or NOX4 siRNA, CLP WT group and CLP groups made in NOX2 or p47phox knockout mice. Freshly prepared lung OCT sections were incubated with DHE (2 μmol/L) in the dark for 30 min, and fluorescent images were captured using a Nikon A1R Confocal Microscope. Photographs were taken at ×200 magnification. Data are presented as Mean±SEM. n = 10. **p < 0.01 vs. CLP group. (B) Representative Western blots and quantitative data of NOX1, NOX2, NOX4 and p22phox protein expression in mice at 1, 2, 4, 6 h post CLP. Data are presented as Mean±SEM; n = 3. *p < 0.05, **p < 0.01 vs. 0 h group. (C) Representative Western blots and quantitative data of NOX1, NOX2, NOX4, and p22phox expression in lung tissues of control siRNA or NOX1 siRNA treated mice with or without CLP for 2 h, indicating that NOX1 siRNA attenuated the expression of NOX1, while having no effects on the expression of NOX2, NOX4, and p22phox. Data are presented as Mean±SEM; n = 3. **p < 0.01. (D) Representative Western blots and quantitative data of NOX1, NOX2, NOX4, and p22phox expression in lung tissues of WT and NOX2 knockout mice with or without CLP for 2 h, indicating that NOX2 knockout had absence of NOX2 but upregulated expression of NOX4 and p22phox. Data are presented as Mean±SEM; n = 3. **p < 0.01. (E) Representative Western blots and quantitative data of NOX1, NOX2, NOX4, and p22phox in lung tissues of control siRNA or NOX4 siRNA treated mice with or without CLP for 2 h, indicating that NOX4 siRNA attenuated the expression of NOX4 and p22phox, while having no effect on the protein expression of NOX1 and NOX2. Data are presented as Mean±SEM; n = 3. **p < 0.01. (F) Representative Western blots and quantitative data of p47phox and p22phox in lung tissues of WT or p47phox knockout mice with or without CLP for 2 h, indicating that p47phox knockout had absence of p47phox but upregulated expression of p22phox. Data are presented as Mean±SEM; n = 3. *p < 0.05.