Figure 1

NOX2-generated ROS is responsible for hepatoma-induced autophagy and M2 macrophage polarization. (a) BMDMs were pretreated with or without N-acetyl cysteine (NAC, 10 mM) for 30 min and then incubated with MCM for another 24 h. The ROS induction was analyzed by flow cytometry. (b) BMDMs were pretreated with or without N-acetyl cysteine (NAC, 10 mM) for 30 min and then incubated with MCM for another 24 h. Supernatants and cell lysates were then collected for determining IL-10 expression by ELISA and CD206 expression by flow cytometry, respectively. (c) BMDMs were treated with MCM in the presence or absence of NAC (1, 5, or 10 mM) for 24 h. Cell lysates were collected to determine the expression of NF-κB p65, LC3-I/II, and p62 by Western blotting. (d) Wild type (WT) or gp91^−/− BMDMs were treated with MCM for the indicated time and the ROS production was determined by flow cytometry. (e) WT or gp91^−/− BMDMs were treated with MCM for 24 h and supernatants were collected to analyze the production of IL-10 by ELISA. (f) NOX2-depedent autophagy is induced by MCM. WT or gp91^−/− BMDMs were treated with MCM for 12 h and the LC3 punctation was observed and quantified by confocal microscopy. (g) WT or gp91^−/− BMDMs were treated with MCM for the indicated time. Their cell lysates were then collected to determine the expression of NF-κB p65, LC3-I/II, and p62 by Western blotting. *p < 0.05; **p < 0.001; ***p < 0.0001.