Fig 5. Blocking ROS reduced ConA-induced p38/IκB phosphorylation and subsequent pro-inflammatory responses in Atg7-silenced macrophages.

A. Raw264.7 cells were pretreated with NAC (1 mM), SB202190 (10 μM) and Sodium 4-aminosalicylate (10 μM), respectively, for 30 min prior to 10 μg/ml ConA treatment for 24 h. Cell viability was measured by an MTT assay and ROS production detected by H₂DCF-DA. B. Western blotting on p-p38 and p-IκB phosphorylation levels in cultured cells. Densitometry of quantification of p-p38 and p-IκB phosphorylation. * p<0.05 and ** p<0.01 vs. controls, # p<0.05 vs. non NAC-treatments. C. ELISA of TNF-α, IFN-γ, IL-6 and IL-1β levels in the supernatant of cultured cells. Raw264.7 cells were transfected with Atg7 siRNA or scrambles siRNA, pretreated with 1 mM NAC for 30 min and then treated with 10 μg/ml ConA for 24 h. Representative results were from 3 independent experiments. Data are presented as mean±SD; * p<0.05 and ** p<0.01 vs. control, # p<0.05 and ## p<0.01 vs. ConA alone.