Fig. 6

ExMt-derived ROS inhibits autophagy and promotes mTOR-dependent macrophage activation in RAW264.7 cells. (A)Protein levels of P62, LC3B and β-actin were expressed in PBS, exMt and exMt + Dynasore treated RAW264.7 cells and related statistical analysis (n = 5/group, one-way ANOVA). (B) Protein levels of P62, LC3B and β-actin were expressed in PBS, fresh exMt and fixed exMt-treated RAW264.7 cells and related statistical analysis (n = 5/group, one-way ANOVA). (C) Protein levels of P62, LC3B and β-actin were expressed in PBS, exMt and exMt + NAC treated RAW264.7 cells and related statistical analysis (n = 5/group, one-way ANOVA). (D) ELISA measurements of pro-inflammatory cytokines in the culture supernatant of PBS, exMt and exMt + NAC treated RAW264.7 cells treated with exMt and rapamycin (n = 4/group, one-way ANOVA). (E) Flow cytometry analysis showing a decrease in CD86-positive cells in PBS, exMt and exMt + NAC treated RAW264.7 cells (n = 3–4/group, one-way ANOVA). (F) Protein levels of P62, LC3B and β-actin were expressed in exMt or exMt + Dynasore treated RAW264.7 and related statistical analysis (n = 5/group, one-way ANOVA). Protein levels of P62, LC3B and β-actin were expressed in exMt or exMT + Rapamycin treated RAW264.7 and related statistical analysis (n = 5/group, one-way ANOVA). (G) Flow cytometry analysis showing a decrease in CD86-positive cells after rapamycin treatment (n = 4/group, one-way ANOVA). (H) ELISA measurements of pro-inflammatory cytokines in the culture supernatant of RAW264.7 cells treated with exMt and rapamycin (n = 4/group, one-way ANOVA)