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 6. CARD6 knockdown-promoted oxidative stress is ROS dependent in LPS-incubated BV2 cells. (AE) BV2 cells transfected with or without siCARD6 were pre-treated with NAC (5 mM) or PLM (10 μM) for 2 h, followed by LPS (100 ng/ml) treatment for another 24 h. Then, all cells were collected for further studies. (A) DCF-DA analysis was used for the calculation of ROS production. Scale bar: 100 μm. (B) SOD activity in cells was measured. (C) RT-qPCR analysis was used to measure the mRNA levels of SOD1, SOD2, HO1, NQO1, GCLM and GCLC in cells. (D) Nrf2 and Keap1 mRNA expression levels were determined by RT-qPCR. (E) Cytosolic Nrf2 and Keap1, and nuclear Nrf2 expression levels were assessed using western blot analysis. Data represented means ± SEM (n=6 each group). *p < 0.05 and **p < 0.01.