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. 2016 Nov 9;11(11):e0165701. doi: 10.1371/journal.pone.0165701

Fig 4. Inhibition of autophagy enhances cytokines production and ROS generation induced by F. nucleatum infection.

Fig 4

(A, B, and C) After pretreatment of 0.1% DMSO, 3-MA (2mM), Baf-A1 (10 nM) or Rapa (100 nM), Caco-2 cells were infected with F. nucleatum (MOI = 100:1) for 12 hrs. Supernatants of medium were assessed by ELISA for levels of IL-8, IL-1β and TNF-α. (D, E and F) Production of IL-8, IL-1β and TNF-α in Caco-2 cells transfected with siRNA specific for ATG5 or ATG12 (50 nM) for 24h and infected with F. nucleatum (MOI = 100) for 12 hrs, as assessed by ELISA. (G) siRNAs targeting ATG5 or ATG12 (100 nM each) transfected Caco-2 cells for 24 hrs, and the protein levels were assayed by Western blotting. (H and I) After pretreatment of 0.1% DMSO, 3-MA (2mM), Baf-A1 (10 nM) or Rapa (100 nM), or transfected with siRNA specific for ATG5 or ATG12 (50 nM) for 24h, Caco-2 cells were infected with F. nucleatum (MOI = 100:1) for 12 hrs. ROS generation was detected by DCFH-DA assay. The data shown are the means ±SEM of three experiments. *, P<0.05.