Figure 7. CARD6 silence-accelerated apoptosis and inflammation are regulated by ROS production in BV2 cells exposed to LPS. (A–D) BV2 cells in the presence or absence of siCARD6 transfection were pre-treated with NAC (5 mM) or PLM (10 μM) for 2 h, and then were subjected to LPS (100 ng/ml) exposure for another 24 h. Subsequently, all cells were harvested for further analysis. (A) TUNEL and Cyto-c staining of BV2 cells. TUNEL- and Cyto-c-positive cells were quantified. Scale bar: 100 μm. (B) Protein expression levels of Cyto-c, Bcl-2, Bax and cleaved Caspase-3 were evaluated by western blot analysis. (C) RT-qPCR analysis of TNF-α and IL-1β in cells. (D) Protein expression levels of phosphorylated NF-κB in whole cells and NF-κB in nuclear were measured by western blot analysis. Data represented means ± SEM (n=6 each group). *p < 0.05 and **p < 0.01.