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 3. CARD6 knockout enhances apoptosis after SCI. (A) Representative images of NeuN/TUNEL double staining in dorsal horn of mice. The number of apoptotic cells (TUNEL positive) was quantified. Scale bar: 100 μm. (B, C) Western blot analysis of Bcl-2, Bax and cleaved Caspase-3 protein expression levels in the lumbar spinal cord segments. (D) Western blot analysis of mitochondrial and whole cell Cyto-c in the lumbar spinal cord segments. (E, F) BV2 cells with or without CARD6 knockdown were treated with LPS (100 ng/ml) for 24 h. (E) Then, IF staining was used to determine Iba1, Cyto-c and TUNEL levels. The quantification of Iba1, Cyto-c and TUNEL expression levels was exhibited. Scale bar: 50 μm. (F) Western blot analysis was used to calculate Cyto-c protein levels in mitochondria, or Cyto-c, Bcl-2, Bax and cleaved Caspase-3 expression in whole cell as indicated. 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.