Research Paper Volume 11, Issue 24 pp 12213—12235

Targeting CARD6 attenuates spinal cord injury (SCI) in mice through inhibiting apoptosis, inflammation and oxidative stress associated ROS production

Figure 5. CARD6 knockout promotes oxidative stress after SCI. Measurements of (A) SOD, CAT, GSH, (B) MDA, H2O2 and 3-NT in spinal cords of mice at 3 days after SCI. (C) Representative images of 4-HNE by IF staining in dorsal horn of mice. The relative expression of 4-HNE was quantified. Scale bar: 100 μm. (D) RT-qPCR analysis of SOD1, SOD2, HO1, NQO1, GCLM and GCLC mRNA levels in the lumbar spinal cord segments. (E) RT-qPCR analysis of Nrf2 and Keap1 mRNA expression levels in the lumbar spinal cord segments. (F) Western blot analysis of cytosolic Nrf2 and Keap1, and nuclear Nrf2 protein expression levels in the lumbar spinal cord segments. (G) Representative images of Nrf2 by IF staining in dorsal horn of mice. The quantification of Nrf2 expression levels was showed. Scale bar: 100 μm. (H) IF staining of Nrf2 in BV2 cells transfected with siCARD6 followed by 24 h of LPS (100 ng/ml) stimulation. Then, Nrf2 expression levels were quantified. Scale bar: 25 μm. Data represented means ± SEM (n=8 each group for in vivo studies; n=6 each group for in vitro studies). *p < 0.05 and **p < 0.01.