FIGURE 3.

Bacillus amyloliquefaciens SC06 (Ba) treatment induces autophagy in RAW264.7 cells (A) RAW264.7 cells were treated as follows. Control: Untreated cells. Ba: Cells were incubated with 10⁸ cfu/ml Ba for 6 h. Rapa: Positive control. Cells were treated with rapamycin (2 μM, 6 h). All cells were then stained using immunofluorescence technique and observed under a confocal microscopy. The scale bar represents 5 μm (B) Statistical analyses of the number of positive cells with > 3 green puncta. Values are from 100 cells per sample (t-test, ^∗∗∗p < 0.001). Data are representative of three individual experiments with similar results. (C) Cells were pretreated with Chloroquine (Cq, 50 μM, 3 h) and then incubated with 10⁸ cfu/ml of Ba for 6 h. The levels of LC3 and p62 were identified by western blotting. (D,E) Analyses of LC3-II/LC3-I or p62/β-actin using Image J (one-way ANOVA; Tukey test, ^∗p < 0.05, ^∗∗p < 0.01). Data are representative of three individual experiments with similar results.